Figure 2. HSP47 can regulate ESC-SMC differentiation.

A and B, knockdown of HSP47 abolishes HSP47 expression and attenuates ESC-SMC differentiation. ESCs were infected with non-coding (NC) or HSP47 shRNA lentiviruses for 24 h prior to SMC differentiation, followed by Western blot analysis (A) and quantification (B). C–E, over-expression of HSP47 induces SMC differentiation of ESC. ESCs were transfected either with a pCMV6-HSP47 or mock plasmid (1 µg/1×10⁶ cells) via electroporation, followed by real time RT-PCR (C), Western blotting (D, E) and Immunofluorescent staining with HSP47 (green), calponin (red) (F) and SM-22α (red) (G) antibodies. DAPI was included to counterstain the nucleus (blue). Scale bar, 30 µm. Images and blots shown are representative of at least three separate experiments, whereas graphs are shown as mean ± SEM of at least three independent experiments, *P<0.05, **P<0.01, ***P<0.005 fold increase compared with mock control, dotted line.