Fig. 3

Silencing the Rac1 reversed the hPTH(1–34)-migration of fibroblasts and HaCaT cells. A–D The expression of Rac1 in siRNA-transfected fibroblasts (A) and HaCaT cells (C) were evaluated by western blot analysis after treated with or without hPTH. The intensity of each band was measured and normalized to GAPDH then calculated as the ratio of the controls (B and D). E–H siRNA-transfected fibroblasts (E) and HaCaT cells (G) were incubated with or without hPTH(1–34) for 12 h and the lamellipodia in two types of cells were stained with rhodamine-phalloidin. The ratio of lamellipodia areas versus total cell areas was measured in fibroblasts (F) and HaCaT cells (H). I and J Representative images of wound scratching assays on siRNA-transfected fibroblasts (I) and HaCaT cells (J) after incubating with or without hPTH(1–34). Images were captured at 0 h and 24 h. K and L Representative images of siRNA-transfected Transwell assay on fibroblasts (K) and HaCaT cells (L) after incubation with / without hPTH(1–34) for 48 h. M and N Quantification and analysis of the migrated areas in hPTH(1–34)-treated fibroblasts (M) and HaCaT cells (N) using imageJ software. O and P Quantification of the migrated fibroblasts (O) and HaCaT cells (P) after subjected to hPTH(1–34) or PBS for 48 h. The data represent mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001 vs control group. #p < 0.05; ##p < 0.01; ###p < 0.001 vs hPTH(1–34) group. Scale bar: 25 μm in (E), 10 μm in (G), 50 μm in (I–L). NC Negative control