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. 2012 Aug 28;7(8):e43272. doi: 10.1371/journal.pone.0043272

Figure 3. TGF-β depletion in MSCs reverses the promotive effect on metastasis and EMT of HCC cells induced by MSCs in inflammation microenvironment.

Figure 3

(A) Expression of EMT genes was detected by qPCR (normalized to β-actin); (B) E-cadherin and Vimentin expression was performed by immunofluorescent staining, nuclei were counterstained with DAPI (×200). (C) Western blot was used to detect the expression of N-cadherin, Twist and β-catenin, SMMC-7721 cells co-cultured with MSCssi-TGFβ stimulated by both IFNγ and TNFα did not present EMT; (D) The wound healing assay was employed to determine the migration of SMMC-7721 cells (×200); (E) Invasiveness of SMMC-7721 cells was determined using Transwell assay; (F) The metastatic liver nodules in nude mice by splenic-vein injection of SMMC-7721 cells. The arrows indicate the metastatic tumor on the surface of the liver (upper). H&E staining was performed on serial sections of metastatic tumors and normal liver (bottom, ×100); (G) The number of nodules were quantified on livers of nude mice (n = 10 per group) 6 weeks after splenic vein injection of SMMC-7721 cells. (*P<0.05 versus Control group; #P<0.05 versus MSCs(IFNγ+TNFα) and MSCs(IFNγ+TNFα)vector; ×200).