10.1371/journal.pone.0057855.g002 Figure 2  Deletion reporter gene analysis of LPS and PMA-responsive regions in the murine SerpinB2 promoter.
(A) Schematic representation of the murine SerpinB2 promoter and 5′ deletion luciferase reporter constructs. The murine 5′ flanking region from −4480 to +92 was inserted upstream of the luciferase reporter gene and 5′ deletions were generated using restriction enzyme sites or with specific oligonucleotide PCR primers. Construct names indicate the most 5′ nucleotide of murine SerpinB2 5′ flanking sequence. (B) RAW 264.7 macrophages were transiently transfected with the indicated murine SerpinB2 promoter-luciferase reporter constructs and control plasmids. Cells were either left untreated or treated with 100 ng/ml LPS or 40 ng/ml PMA for 16 hrs. Shown is the relative luciferase reporter gene activity following treatment. The results represent the mean and SEM of four independent experiments. (C) DNA sequence conservation between the human and murine SerpinB2 5′ flanking regions. Schematic representation of the human and murine SerpinB2 5′ flanking regions with regions of nucleotide sequence identity indicated by the same colored boxes. Homologous regions are interrupted by repetitive sequence elements in both 5′ flanking regions. Alu = Alu repeat, ID4 =  ID4 short interspersed nuclear repeat (SINE), L1 =  L1 long interspersed nuclear element (LINE), L2 =  L2 LINE, MIR = MIR SINE, MLT1L = MLT1L long terminal repeat (LTR), (TATG)n = TATG tetranucleotide repeat, tis = transcription initiation site.