Fig. 5

HBx promotes ubiquitination and autophagy degradation of cGAS. (a–b) SMMC-7721 cells were transfected with an empty vector or HBx-Flag plasmid. At 12 h posttransfection, treated with Rapamycin or 3-MA or MG132 (10 mM) for 12 h, dimethyl sulfoxide (DMSO) was used for controls. Cells were harvested and subjected to WB analysis. (c, d) SMMC-7721 cells were transfected with pGFP-LC3 and empty vector or HBx- Flag plasmid. At 12 h posttransfection, treated with Rapamycin or 3MA or MG132 (10 mM) for 12 h, dimethyl sulfoxide (DMSO) was used for controls. Moreover, autophagic dots were observed by a fluorescence microscope. Quantitation of the autophagic cells in SMMC-7721. Cells with more than three fluorescent particles are considered autophagy-activated cells. Meanwhile, the cells were harvested and subjected to WB analysis. (e) SMMC-7721 cells were transfected with an empty vector or HBx- Flag plasmid. At 12 h posttransfection, cells were harvested and subjected to WB analysis. (f–h) L02 were transfected with ISD, empty vector or HBV1.3 plasmid, IFNβ, ISG54, and ISG 56 were detected by qRT-PCR analysis at 12 h posttransfection, HBV core protein was detected by WB analysis. (i–k) HepG2 cells and HepG2.2.15 cells were transfected with ISD, IFNβ, ISG54, and ISG 56 were detected by qRT-PCR analysis at 12 h posttransfection, HBV core protein was detected by WB analysis