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Mediates both Constitutive and LPS-induced SerpinB2 mRNA Expression in MEFs and Inflammatory Primary Macrophages\nTo investigate the importance of C/EBP-β to endogenous SerpinB2 mRNA expression in response to LPS, we utilized C/EBP-β-null (Cebpb −/−) and wild-type MEFs (Cebpb+/+), since RAW264.7 cells constitutively express endogenous C/EBP-β. Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

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Mediates both Constitutive and LPS-induced SerpinB2 mRNA Expression in MEFs and Inflammatory Primary Macrophages\nTo investigate the importance of C/EBP-β to endogenous SerpinB2 mRNA expression in response to LPS, we utilized C/EBP-β-null (Cebpb −/−) and wild-type MEFs (Cebpb+/+), since RAW264.7 cells constitutively express endogenous C/EBP-β. Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

{"project":"bionlp-st-ge-2016-test-proteins","denotations":[{"id":"T24616","span":{"begin":2195,"end":2203},"obj":"Protein"},{"id":"T24615","span":{"begin":2085,"end":2092},"obj":"Protein"},{"id":"T24614","span":{"begin":1914,"end":1919},"obj":"Protein"},{"id":"T24613","span":{"begin":1659,"end":1666},"obj":"Protein"},{"id":"T24612","span":{"begin":1623,"end":1631},"obj":"Protein"},{"id":"T24611","span":{"begin":1526,"end":1531},"obj":"Protein"},{"id":"T24610","span":{"begin":1513,"end":1518},"obj":"Protein"},{"id":"T24609","span":{"begin":1475,"end":1483},"obj":"Protein"},{"id":"T24608","span":{"begin":1401,"end":1406},"obj":"Protein"},{"id":"T24607","span":{"begin":1375,"end":1380},"obj":"Protein"},{"id":"T24606","span":{"begin":1334,"end":1342},"obj":"Protein"},{"id":"T24605","span":{"begin":1293,"end":1301},"obj":"Protein"},{"id":"T24604","span":{"begin":1239,"end":1246},"obj":"Protein"},{"id":"T14036","span":{"begin":1180,"end":1188},"obj":"Protein"},{"id":"T14035","span":{"begin":1087,"end":1094},"obj":"Protein"},{"id":"T14034","span":{"begin":1005,"end":1012},"obj":"Protein"},{"id":"T14033","span":{"begin":949,"end":957},"obj":"Protein"},{"id":"T14032","span":{"begin":796,"end":803},"obj":"Protein"},{"id":"T14031","span":{"begin":647,"end":652},"obj":"Protein"},{"id":"T14030","span":{"begin":592,"end":600},"obj":"Protein"},{"id":"T14029","span":{"begin":515,"end":523},"obj":"Protein"},{"id":"T14028","span":{"begin":459,"end":467},"obj":"Protein"},{"id":"T14027","span":{"begin":429,"end":436},"obj":"Protein"},{"id":"T14026","span":{"begin":401,"end":409},"obj":"Protein"},{"id":"T14025","span":{"begin":344,"end":351},"obj":"Protein"},{"id":"T14024","span":{"begin":278,"end":283},"obj":"Protein"},{"id":"T14023","span":{"begin":247,"end":252},"obj":"Protein"},{"id":"T14022","span":{"begin":233,"end":240},"obj":"Protein"},{"id":"T14021","span":{"begin":176,"end":184},"obj":"Protein"},{"id":"T14020","span":{"begin":154,"end":161},"obj":"Protein"},{"id":"T14019","span":{"begin":51,"end":59},"obj":"Protein"},{"id":"T14018","span":{"begin":0,"end":7},"obj":"Protein"}],"namespaces":[{"prefix":"_base","uri":"http://bionlp.dbcls.jp/ontology/ge.owl#"}],"text":"C/EBP-β Mediates both Constitutive and LPS-induced SerpinB2 mRNA Expression in MEFs and Inflammatory Primary Macrophages\nTo investigate the importance of C/EBP-β to endogenous SerpinB2 mRNA expression in response to LPS, we utilized C/EBP-β-null (Cebpb −/−) and wild-type MEFs (Cebpb+/+), since RAW264.7 cells constitutively express endogenous C/EBP-β. Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

{"project":"bionlp-st-ge-2016-uniprot","denotations":[{"id":"T24865","span":{"begin":2195,"end":2203},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T24864","span":{"begin":1623,"end":1631},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T24863","span":{"begin":1475,"end":1483},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14300","span":{"begin":1005,"end":1012},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T14299","span":{"begin":796,"end":803},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T14298","span":{"begin":429,"end":436},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T14297","span":{"begin":344,"end":351},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T14296","span":{"begin":233,"end":240},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T14295","span":{"begin":154,"end":161},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T14294","span":{"begin":0,"end":7},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T14309","span":{"begin":1180,"end":1188},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14308","span":{"begin":949,"end":957},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14307","span":{"begin":592,"end":600},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14306","span":{"begin":515,"end":523},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14305","span":{"begin":459,"end":467},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14304","span":{"begin":401,"end":409},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14303","span":{"begin":176,"end":184},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14302","span":{"begin":51,"end":59},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T14301","span":{"begin":1087,"end":1094},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T24854","span":{"begin":1659,"end":1666},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T24853","span":{"begin":1239,"end":1246},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T24862","span":{"begin":1334,"end":1342},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T24861","span":{"begin":1293,"end":1301},"obj":"http://www.uniprot.org/uniprot/P05120"},{"id":"T24860","span":{"begin":2125,"end":2130},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T24859","span":{"begin":1914,"end":1919},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T24858","span":{"begin":1526,"end":1531},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T24857","span":{"begin":1513,"end":1518},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T24856","span":{"begin":1401,"end":1406},"obj":"http://www.uniprot.org/uniprot/P28033"},{"id":"T24855","span":{"begin":1375,"end":1380},"obj":"http://www.uniprot.org/uniprot/P28033"}],"namespaces":[{"prefix":"_base","uri":"http://www.uniprot.org/uniprot/"}],"text":"C/EBP-β Mediates both Constitutive and LPS-induced SerpinB2 mRNA Expression in MEFs and Inflammatory Primary Macrophages\nTo investigate the importance of C/EBP-β to endogenous SerpinB2 mRNA expression in response to LPS, we utilized C/EBP-β-null (Cebpb −/−) and wild-type MEFs (Cebpb+/+), since RAW264.7 cells constitutively express endogenous C/EBP-β. Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

{"project":"GO-BP","denotations":[{"id":"T14038","span":{"begin":1152,"end":1165},"obj":"http://purl.obolibrary.org/obo/GO_0006351"},{"id":"T14037","span":{"begin":204,"end":219},"obj":"http://purl.obolibrary.org/obo/GO_0032496"}],"text":"C/EBP-β Mediates both Constitutive and LPS-induced SerpinB2 mRNA Expression in MEFs and Inflammatory Primary Macrophages\nTo investigate the importance of C/EBP-β to endogenous SerpinB2 mRNA expression in response to LPS, we utilized C/EBP-β-null (Cebpb −/−) and wild-type MEFs (Cebpb+/+), since RAW264.7 cells constitutively express endogenous C/EBP-β. Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

{"project":"GO-CC","denotations":[{"id":"T14039","span":{"begin":304,"end":309},"obj":"http://purl.obolibrary.org/obo/GO_0005623"}],"text":"C/EBP-β Mediates both Constitutive and LPS-induced SerpinB2 mRNA Expression in MEFs and Inflammatory Primary Macrophages\nTo investigate the importance of C/EBP-β to endogenous SerpinB2 mRNA expression in response to LPS, we utilized C/EBP-β-null (Cebpb −/−) and wild-type MEFs (Cebpb+/+), since RAW264.7 cells constitutively express endogenous C/EBP-β. Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

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Mediates both Constitutive and LPS-induced SerpinB2 mRNA Expression in MEFs and Inflammatory Primary Macrophages\nTo investigate the importance of C/EBP-β to endogenous SerpinB2 mRNA expression in response to LPS, we utilized C/EBP-β-null (Cebpb −/−) and wild-type MEFs (Cebpb+/+), since RAW264.7 cells constitutively express endogenous C/EBP-β. Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

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Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

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Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

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Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

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Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

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Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}

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Wild-type MEFs express low levels of endogenous SerpinB2 and the absence of C/EBP-β attenuated endogenous SerpinB2 mRNA expression. LPS stimulated an increase in SerpinB2 mRNA expression in wild-type MEFs (Fig. 6A), whereas LPS-stimulated SerpinB2 mRNA expression was significantly dampened in Cebpb−/− MEFs as compared to wild-type. We next tested if similar effects could be seen in thioglycollate-elicited inflammatory macrophages in which C/EBP-β expression was knocked down using species-specific lentiviral shRNAs (Fig. 6B, left). As was observed in MEFs, both constitutive and LPS-induced SerpinB2 mRNA expression was significantly decreased in C/EBP-β-deficient inflammatory macrophages (Fig. 6B, right). These data show that C/EBP-β is critical for mediating constitutive and LPS-inducible transcription of endogenous SerpinB2 mRNA.\n10.1371/journal.pone.0057855.g006 Figure 6 C/EBP-β is essential for constitutive and LPS-induced SerpinB2 mRNA expression.\n(A) Endogenous SerpinB2 mRNA expression is abrogated in Cebpb−/− MEFs compared to Cebpb+/+ MEFs in the absence and presence of LPS. qPCR analysis of murine SerpinB2 mRNA expression in untreated Cebpb+/+ and Cebpb−/− MEFs, and after simulation with LPS (100 ng/ml) for the indicated times. (B) Endogenous SerpinB2 expression is abrogated in C/EBP-β-deficient inflammatory macrophages. Thioglycollate-elicited peritoneal macrophages (TG macs) were infected with human and murine specific lentiviral shRNAs. Human CEBPB shRNA serves as the non-silencing control since it does not target the murine Cebpb sequence [38]. Lentiviral transduced macrophages were stimulated with LPS (100 ng/ml) for 4 hrs. Left: Western blot analysis shows effective knockdown of endogenous C/EBP-β following infection with murine Cebpb shRNA and not human CEBPB shRNA. Right: qPCR analysis of murine SerpinB2 mRNA expression in the lentiviral transduced peritoneal macrophages. The results represent the mean and SEM of two independent experiments performed in duplicate or triplicate. (*, p\u003c0.05, two-way ANOVA)."}