Figure 3
Demethylation correlated with the extracellular matrix production of primary lung fibroblasts.
Extracellular matrix production of primary normal and IPF fibroblasts treated with 5′-azacytidine at 5 μM or control medium for 24 hours or 48 hours was assessed with Western, RT-PCR and ELISA. Col1 protein levels in normal (a) and IPF (b) lung fibroblasts were analyzed with Western blotting. β-actin was used as a loading control (normal n = 3 and IPF n = 3). Col1 mRNA levels in normal (c) and IPF (d) lung fibroblasts were assessed with RT-PCR. GAPDH was used as a loading control. (Col1, Collagen 1, C, control, normal n = 4 and IPF n = 6, *P < 0.05; **P < 0.01; ***P < 0,001; data are mean and SEM, unpaired t test) (e). Has2 mRNA expression in primary normal and IPF fibroblasts treated with 5′-azacytidine were assessed with RT-PCR (C, control, normal n = 8 and IPF n = 7. *P < 0.05, paired t test). (f) The levels of HA in the supernatant of normal and IPF lung fibroblasts treated with 5′-azacytidine at 5 μM for 24 hours (C, control, HA, Hyaluronan, Normal n = 8 and IPF n = 7, *P < 0.05; **P < 0.01, paired t test). The full size blots were shown in the Supplementary Fig. S3.
