Fig. 5
Activated fibroblasts by miR-1247-3p promote liver cancer progression. a IL-6 and IL-8 secretion from MRC5 expressed miR-1247-3p or control were detected by ELISA assays. b qRT-PCR analysis of stemness-associated genes expression in SMMC-7721 with indicated treatments. c Spheroid formation ability of SMMC-7721 treated with indicated CM. Representative images were shown and spheroid were counted. Scale bar, 150 μm. d Relative spheroid formation ability of SMMC-7721 treated with indicated CM containing anti-IL-6/anti-IL-8 antibody or IgG control antibody. Representative images were shown. Scale bar, 150 μm. e qRT-PCR analysis of EMT-associated genes expression in SMMC-7721 with indicated treatments. f Migration assay of SMMC-7721 treated with indicated CM. Representative images were shown and migrated cells were counted. Scale bar, 150 μm. g Relative migration ability of SMMC-7721 treated with indicated CM containing anti-IL-6/anti-IL-8 antibody or IgG control antibody. Representative images were shown. Scale bar, 150 μm. h CCK8 assay of SMMC-7721 treated with indicated CM containing anti-IL-6/anti-IL-8 antibody or IgG control antibody in the presence of sorafenib. i Relative cell viabilities of SMMC-7721 treated with indicated CM containing anti-IL-6/anti-IL-8 antibody or IgG control antibody in presence of sorafenib. j Western blotting assay of indicated proteins in SMMC-7721 with indicated treatments. k Xenograft assays of SMMC-7721 with indicated treatments were performed on nude mice. Representative tumors (left) and tumors growth curves (right) were shown. l ELISA assay of IL-6 and IL-8 secretion from NF#1 or CAF#1 isolated from normal or primary HCC tissues. m B4GALT3 expression level was determined in isolated NFs and CAFs by qRT-PCR analysis. Each experiment was performed in triplicate and data are presented as mean ± s.d. Student’s t-test was used to analyze the data (*p < 0.05; **p < 0.01; ***p < 0.001)
