Fig. 2

CAFs enhanced the ability of movement, migration and invasion of Hep2 cells via the high expression of ROCK1. (A) ROCK1 IF staining in CAFs/si-NC or CAFs/si-ROCK1 cells. ROCK1 level was reduced in Hep2/CAFs/si-ROCK1 cells. (D). The inhibition effect and CAFs associative phenotype were verified by Western Blot. ROCK1, FAP, α-SMA, FSP1, NG2 and PDGF-β levels were significantly decreased in Hep2/CAFs/si-ROCK1 cells. (E). Protein ratio in CAFs/si-NC and NFs/ROCK1 cells (*P < 0.05, **P < 0.01). (B) Hep2/CAFs/si-NC or Hep2/CAFs/si-ROCK1 cells on cell mobility as assessed via wound healing assay. Hep2/CAFs/si-ROCK1 cells decreased the mobility. (C) Wound size in Hep2/CAFs/si-NC or Hep2/CAFs/si-ROCK1 cells (*P < 0.05). (F). Hep2 cells co-cultured with CAFs/si-NC or CAFs/si-ROCK1 on cell migration and invasion as measured via trans-well assay. Hep2/CAFs/si-ROCK1 inhibit the ability of migration and invasion. (G). Cell number in every field in Hep2/NFs/vector and Hep2/CAFs/si-ROCK1 cells (*P < 0.05, **P < 0.01). (H). Plasmid transfection was used to up-regulate ROCK1 in NFs, the effect of transfection was showed via IF. (K). The expression of ROCK1 and CAFs associative phenotype were verified by Western Blot. ROCK1, FAP, α-SMA, FSP1, NG2 and PDGF-β levels were significantly increased. (L). Protein ratio in NFs/vector or NFs/ROCK1 cells (*P < 0.05, **P < 0.01). (I). Hep2/NFs/vector or Hep2/NFs/ROCK1 cells on cell mobility as assessed via wound healing assay. Hep2/NFs/ROCK1 cells increased the mobility. (J). Wound size in Hep2/CAFs or Hep2/NFs cells (**P < 0.01). (M). Hep2/NFs/vector or Hep2/NFs/ROCK1 cells on cell migration and invasion as measured via trans-well assay. Hep2/NFs/ROCK1 cells stimulated the ability of migration and invasion. (N). Cell number in every field in Hep2/NFs/vector or Hep2/NFs/ROCK1 cells (*P < 0.05, **P < 0.01)