Figure 2
Demethylation decreased BMPER expression in normal and IPF fibroblasts and regulated the BMPER promoter activity.
Primary normal (a,b) and IPF lung fibroblasts (c,d) were treated with 5′-azacytidine at 5 μM for 24 hours. BMPER protein levels in normal and IPF lung fibroblasts were analyzed with Western blotting (4 samples are shown). β-actin was used as a loading control. The relative amount was shown with densitometry analysis (normal n = 7 and IPF n = 5; N, normal; C, control; 5′aza, 5′-azacytidine; *P < 0.05; paired t test). (e) BMPER mRNA expression was assessed in human normal and IPF lung fibroblasts after 5 μM 5′-azacytidine treatment for 24 hours with qRT-PCR (normal n = 5 and IPF n = 7; C, control; 5′aza, 5′-azacytidine; *P < 0.05, paired t test). (f) Luciferase activity of human BMPER promoter was measured in HEK293 cells treated with 5′-azacytidine at 5 μM for 24 hours (***P < 0.001, data are mean and SEM, unpaired t test). (g) Cell viability of human lung fibroblasts with and without 5′-azacytidine treatment was measured with MTT assay. (n = 5, P > 0.05, paired t test). The full size blots were shown in the Supplementary Fig. S2.
