Fig. 1

CAFs were isolated from LSCC and CAFs enhanced the ability of metastasis of LSCC in vitro and vivo and ROCK1 was highly expressed in CAFs. (A) CAFs specific markers (FAP, α-SMA, FSP1, NG2 and PDGF-β) were determined by Western Blot. The expressions of markers were significantly elevated in CAFs. (B) Protein ratio in NFs, CPFs and CAFs (**P < 0.01). (C) Co-cultured model was used to separate Hep2 cells line and CAFs or NFs. (D) Hep2 cells co-cultured with CAFs or NFs on cell mobility as assessed via wound healing assay. Hep2/CAFs cells increased the mobility. (E) Wound size in Hep2/CAFs or NFs cells (**P < 0.01). (F) Hep2/CAFs or NFs cells on cell migration and invasion as measured via trans-well assay. Hep2/CAFs cells stimulated the ability of migration and invasion. (G) Cells number in every field in Hep2/CAFs or NFs cells (*P < 0.05, **P < 0.01). (H) Hep2/CAFs or NFs cells were inoculated into nude mice and pulmonary nodules were observed after six weeks (N = 5/group). H&E stains of pulmonary nodules (100×), Hep2/CAFs cells demonstrated larger and more frequently lung metastases as compared to Hep2/NFs cells. (I) Pulmonary tissue and nodules were quantified by H&E staining from co-cultured with CAFs or NFs (*P < 0.05). (J) The mRNA expressions of ROCK1 were determined by Rt-PCR. (K). The protein expression of ROCK1 were determined by Western Blot. （L). Protein ratio in NFs, CPFs and CAFs (**P < 0.01)