Fig. 6

Linc00261 blocks TGF-β1 signaling via inhibiting SMAD3 expression and phosphorylation. Western blotting analysis of SMAD3 and p-SMAD3 protein after linc00261 knockdown (A; Huh7 and MHCC-LM3) or linc00261 overexpression (B; SMMC-7721). C Linc00261 overexpression promotes the ubiquitination dependent degradation of SMAD3 reflected by CHX (10 μM) and MG132 (10 μM) treatment in SMMC-7721 cells. D Western blotting analysis of total or p-SMAD3 protein expression in linc00261 overexpression or control cells treated with or without TGF-β1 in SMMC-7721 cells. E Immunofluorescence staining of SMAD3 and p-SMAD3 protein after linc00261 overexpression in SMMC-7721 cells. F IHC analysis of SMAD3 and p-SMAD3 protein levels in vivo. The histograms represent the evaluation of gray values of these western blotting bands. * P < 0.05; ** P < 0.01; *** P < 0.001