Fig. 3: TNFAIP8 induces autophagy in liver cancer cells.

a Fifty micrograms of lysates from HCC cells immunoblotted with indicated antibodies. b Fifty micrograms liver cancer tissue lysates with different stages of liver cancer were immunoblotted with indicated antibodies. c HepG2, SK-Hep1 cells were transfected with EV or TNFAIP8-Myc plasmid for 30 h, and cell lysates from transient and stable HepG2-TNFAIP8 expressing cells were immunoblotted with indicated antibodies. d HepG2 cells were grown in complete medium or serum-free (S.F.) medium for 24 h and then transfected with EV or TNFAIP8-Myc plasmid for 30 h. Fifty micrograms of proteins were immunoblotted. e HepG2 cells were transfected with control siRNA, or TNFAIP8 siRNA (100 nM) for 30 h and lysates were immunoblotted with autophagy related bio-markers. f HepG2 cells were grown in complete medium or serum-free (S.F.) medium for 24 h and then transfected with control siRNA or TNFAIP8 siRNA (100 nM) for 30 h as indicated, and lysates were immunoblotted with indicated antibodies. g HepG2 cells were pretreated with the autophagy inhibitor, 3-MA for 18 h and transfected with EV and TNFAIP8-Myc plasmid for 24 h in the presence of 3-MA and cell lysates were immunoblotted with indicated antibodies. h, i Effect of TNFAIP8 overexpression and 3-MA on LC3β related puncta formation in HepG2 cells were analyzed by Cyto-ID green fluorescence staining (h), and number of puncta were measured from TNFAIP8-transfected or 3-MA pretreated and TNFAIP8-transfected cells and plotted (i). ***P < 0.001 relative to TNFAIP8 transfected. EV: empty vector, Comp. medium: complete medium, S.F. medium: serum-free medium, N: normal, T: tumor. Arrow indicates ATG3 specific band.