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7 The E2f3a Isoform Drives the Differentiation Defect in Rb KO SACs\n(A) Schematic diagrams of the mouse WT, E2f3a−/−, and the Cre-recombined floxed E2f3 loci (indicated here as E2f3−/− for simplicity). E2f3a−/− mice lack most of E2f3 exon 1a and part of intron 1a (red dotted box). Arrows indicate PCR primers. Genotyping of an E2f3a+/− mouse is shown on the right.\n(B) RT-PCR detection of E2f3a and E2f3b mRNA in the retina. The sequences of primers are 1aF (5′-GCCTCTACACCACGCCACAAG-3′), 1bF (5′-CGGAAATGCCCTTACAGC-3′), and 4R (5′-CTCAGTCACTTCTTTGGACAG-3′). WT retina expresses both E2f3a and E2f3b mRNA. As expected, E2f3a−/− retina lacks E2f3a mRNA and still expresses E2f3b mRNA. E2f3−/− retina lacks full-length E2f3a and E2f3b mRNAs, and instead expresses a truncated mRNA lacking exon 3.\n(C) Real-time RT-PCR analysis of E2f genes in P8 retinas of the indicated genotypes. Error bars represent SD of measurements from three animals, and asterisks indicate a significant difference between WT and the indicated genotypes (*, p \u003c0.05; **; p \u003c0.01; ANOVA and Tukey HSD test).\n(D) Rescue of Rb KO SACs by E2f3a deletion. Horizontal retinal sections of the indicated genotypes and ages were stained for nuclei (DAPI, blue), M-phase (PH3, green), and the SAC marker Slc18a3 (red). E2f3a deletion does not suppress ectopic division, but rescues the SAC defect. Scale bars are 50 μm.\nM, molecular size marker."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T37834","span":{"begin":14,"end":19},"obj":"PR_EXT:000028775"},{"id":"T37835","span":{"begin":20,"end":27},"obj":"SO_EXT:0001060"},{"id":"T37836","span":{"begin":39,"end":54},"obj":"GO_RO_EXT:developmental_differentiation_process"},{"id":"T37837","span":{"begin":65,"end":67},"obj":"PR_EXT:000013773"},{"id":"T37838","span":{"begin":106,"end":111},"obj":"NCBITaxon:10088"},{"id":"T37839","span":{"begin":112,"end":114},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T37840","span":{"begin":116,"end":121},"obj":"PR_EXT:000028775"},{"id":"T37841","span":{"begin":121,"end":122},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37842","span":{"begin":123,"end":124},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37843","span":{"begin":138,"end":148},"obj":"GO_SO_EXT:sequence_rearrangement_process"},{"id":"T37844","span":{"begin":149,"end":155},"obj":"SO:0000359"},{"id":"T37845","span":{"begin":156,"end":160},"obj":"PR_EXT:000006854"},{"id":"T37846","span":{"begin":185,"end":189},"obj":"PR_EXT:000006854"},{"id":"T37847","span":{"begin":189,"end":190},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37848","span":{"begin":191,"end":192},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37849","span":{"begin":210,"end":215},"obj":"PR_EXT:000028775"},{"id":"T37850","span":{"begin":215,"end":216},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37851","span":{"begin":217,"end":218},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37852","span":{"begin":219,"end":223},"obj":"NCBITaxon:10088"},{"id":"T37853","span":{"begin":237,"end":241},"obj":"PR_EXT:000006854"},{"id":"T37854","span":{"begin":242,"end":246},"obj":"SO_EXT:0000147"},{"id":"T37855","span":{"begin":262,"end":268},"obj":"SO_EXT:0000188"},{"id":"T37856","span":{"begin":310,"end":317},"obj":"SO_EXT:0000112"},{"id":"T37857","span":{"begin":336,"end":341},"obj":"PR_EXT:000028775"},{"id":"T37858","span":{"begin":341,"end":342},"obj":"SO_EXT:normal_or_wild_type_or_present"},{"id":"T37859","span":{"begin":343,"end":344},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37860","span":{"begin":345,"end":350},"obj":"NCBITaxon:10088"},{"id":"T37861","span":{"begin":378,"end":380},"obj":"GO_EXT:reverse_transcription_or_reverse_transcriptase"},{"id":"T37862","span":{"begin":398,"end":403},"obj":"PR_EXT:000028775"},{"id":"T37863","span":{"begin":414,"end":418},"obj":"CHEBI_SO_EXT:mRNA"},{"id":"T37864","span":{"begin":426,"end":432},"obj":"UBERON:0000966"},{"id":"T37865","span":{"begin":438,"end":447},"obj":"SO_EXT:biological_sequence"},{"id":"T37866","span":{"begin":451,"end":458},"obj":"SO_EXT:0000112"},{"id":"T37867","span":{"begin":568,"end":570},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T37868","span":{"begin":571,"end":577},"obj":"UBERON:0000966"},{"id":"T37869","span":{"begin":578,"end":587},"obj":"GO:0010467"},{"id":"T37870","span":{"begin":593,"end":598},"obj":"PR_EXT:000028775"},{"id":"T37871","span":{"begin":609,"end":613},"obj":"CHEBI_SO_EXT:mRNA"},{"id":"T37872","span":{"begin":628,"end":633},"obj":"PR_EXT:000028775"},{"id":"T37873","span":{"begin":633,"end":634},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37874","span":{"begin":635,"end":636},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37875","span":{"begin":637,"end":643},"obj":"UBERON:0000966"},{"id":"T37876","span":{"begin":650,"end":655},"obj":"PR_EXT:000028775"},{"id":"T37877","span":{"begin":656,"end":660},"obj":"CHEBI_SO_EXT:mRNA"},{"id":"T37878","span":{"begin":671,"end":680},"obj":"GO:0010467"},{"id":"T37879","span":{"begin":687,"end":691},"obj":"CHEBI_SO_EXT:mRNA"},{"id":"T37880","span":{"begin":693,"end":697},"obj":"PR_EXT:000006854"},{"id":"T37881","span":{"begin":697,"end":698},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37882","span":{"begin":699,"end":700},"obj":"SO_EXT:sequence_nullness_or_absence"},{"id":"T37883","span":{"begin":701,"end":707},"obj":"UBERON:0000966"},{"id":"T37884","span":{"begin":726,"end":731},"obj":"PR_EXT:000028775"},{"id":"T37885","span":{"begin":742,"end":747},"obj":"CHEBI_SO_EXT:mRNA"},{"id":"T37886","span":{"begin":761,"end":770},"obj":"GO:0010467"},{"id":"T37887","span":{"begin":773,"end":782},"obj":"SO_EXT:sequence_truncation_process"},{"id":"T37888","span":{"begin":783,"end":787},"obj":"CHEBI_SO_EXT:mRNA"},{"id":"T37889","span":{"begin":796,"end":800},"obj":"SO_EXT:0000147"},{"id":"T37890","span":{"begin":818,"end":820},"obj":"GO_EXT:reverse_transcription_or_reverse_transcriptase"},{"id":"T37891","span":{"begin":837,"end":840},"obj":"PR_EXT:E2F"},{"id":"T37892","span":{"begin":841,"end":846},"obj":"SO_EXT:0000704"},{"id":"T37893","span":{"begin":853,"end":860},"obj":"UBERON:0000966"},{"id":"T37894","span":{"begin":878,"end":887},"obj":"SO_EXT:genotype_or_entity_with_genotype"},{"id":"T37895","span":{"begin":940,"end":947},"obj":"NCBITaxon:33208"},{"id":"T37896","span":{"begin":1005,"end":1007},"obj":"SO_EXT:wild_type_entity_or_quality"},{"id":"T3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7 The E2f3a Isoform Drives the Differentiation Defect in Rb KO SACs\n(A) Schematic diagrams of the mouse WT, E2f3a−/−, and the Cre-recombined floxed E2f3 loci (indicated here as E2f3−/− for simplicity). E2f3a−/− mice lack most of E2f3 exon 1a and part of intron 1a (red dotted box). Arrows indicate PCR primers. Genotyping of an E2f3a+/− mouse is shown on the right.\n(B) RT-PCR detection of E2f3a and E2f3b mRNA in the retina. The sequences of primers are 1aF (5′-GCCTCTACACCACGCCACAAG-3′), 1bF (5′-CGGAAATGCCCTTACAGC-3′), and 4R (5′-CTCAGTCACTTCTTTGGACAG-3′). WT retina expresses both E2f3a and E2f3b mRNA. As expected, E2f3a−/− retina lacks E2f3a mRNA and still expresses E2f3b mRNA. E2f3−/− retina lacks full-length E2f3a and E2f3b mRNAs, and instead expresses a truncated mRNA lacking exon 3.\n(C) Real-time RT-PCR analysis of E2f genes in P8 retinas of the indicated genotypes. Error bars represent SD of measurements from three animals, and asterisks indicate a significant difference between WT and the indicated genotypes (*, p \u003c0.05; **; p \u003c0.01; ANOVA and Tukey HSD test).\n(D) Rescue of Rb KO SACs by E2f3a deletion. Horizontal retinal sections of the indicated genotypes and ages were stained for nuclei (DAPI, blue), M-phase (PH3, green), and the SAC marker Slc18a3 (red). E2f3a deletion does not suppress ectopic division, but rescues the SAC defect. Scale bars are 50 μm.\nM, molecular size marker."}

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