Figure 3

Specific inhibition of CLIC4 expression stops myofibroblast transdifferentiation to fibroblast. MRC-5 cells were transfected with specific CLIC4 siRNA and then treated with or without CM^SKOV3 and TGF-β1. Mock transfection with transfection reagent alone and scrambled siRNA (neg siRNA) served as negative controls. The relative levels of CLIC4 (A), α-SMA (B) transcripts and α-SMA protein expression were assessed by real time RT-PCR, Western blot (C) and immunofluorescent staining (D). Scale bar, 50 μm. Quantitative RT-PCR was used to analyse transcription levels of pro-angiogenic factors, VEGF (E), HGF (F) and α-SMA (G) in MRC-5 cells treated with chloride channel inhibitor IAA-94, NAC or CLIC4 siRNA before addition of CM^SKOV3 and TGF-β1. * P < 0.05.