Fig. 5
Knockdown of CCN2 attenuated the pro-fibrotic functions of fibroblasts by downregulating TGFBR/SMAD signalling pathway. a, b Expression of CCN2 with siCCN2 or with siNC (non-target control siRNA) assessed by RT-qPCR and western blotting (n = 3). c–h Expression of CCN2 and other activation markers (αSMA and COL1) after culturing with TGF-β (4 ng/mL) in transfected fibroblasts analyzed by RT-qPCR and western blotting (n = 3). i Proliferations of the transfected cells measured using the CCK-8 assay at different time intervals (n = 3). j Cell migration evaluated by wound-healing assays (n = 3). Representative images (original magnification, ×40) are shown on the left. Bar = 200 μm. The migration area rates in multiple time intervals are presented on the right. k, l Expression of TGFBR1 and SMAD3 after culturing with TGF-β (4 ng/mL) in transfected fibroblasts analyzed by RT-qPCR (n = 3). m Left: Representative immunofluorescence images of TGFBR1 and SMAD3 in transfected fibroblasts after TGF-β (4 ng/mL) stimulation. Right: Bar plots showing relative fluorescence intensity of TGFBR1 and SMAD3. OD, optical density; *P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.000 1. Data are shown as mean ± SD
