Fig. 3

Knockdown of TROY inhibits stemness properties of HCC cells. (A, B) Western blotting of TROY (A), NANOG, SOX2, and OCT3/4 (B) in HepG2 and Huh7 HCC cells transfected with lentiviruses expressing control shRNA (shNTC) or two independent shRNAs targeting TROY (shb and shc). β-Actin was used as a loading control. (C) Spheroid formation assay (left) and summaries (right) of spheroid numbers in HepG2 and huh7 cells transfected with shNTC or TROY shRNAs. (D) XTT assay reveals a stronger chemoresistance ability of cells with higher TROY expression compared with that of lower TROY expression. Cells were treated with the indicated concentration of Sorafenib (SR) for 48 h. (E) Representative flow cytometry plots (left) and summary (right) of apoptotic cells in HepG2 and huh7 cells transfected with shNTC or TROY shRNAs. Cells were treated with Sorafenib (20 μm) for 24 h. (F) In vivo subcutaneous implantation NOD/SCID mice model induced by HepG2 cells transfected with shNTC or TROY shRNAs. (G) In vivo subcutaneous implantation nude mice model induced by huh7 cells with shNTC or TROY shRNAs. Statistical significances: *, P < 0.05; **, P < 0.01; ***, P < 0.001