HBV expression positively regulates expression of endogenous NSUN2.

(A) Western blot (left) and gray degree analysis (right) of endogenous NSUN2 in HepG2 after pHBV1.3 gradient transfection. Relative levels of NSUN2 protein to GAPDH were quantified using Quantity One. At 48 hours after pHBV1.3 transfection, cells were harvested for Western blot. (B) Western blot of endogenous NSUN2 in HepG2 after transfection with pHBV1.3 and its mutant. Relative levels of NSUN2 protein were quantified using Quantity One. (C) Western blot (left) of endogenous NSUN2 after HBV infection in PHHs with wild-type or mutated virus. At 9 days after HBV infection, cells were harvested for Western blot. The relative levels of NSUN2 protein were quantified using Quantity One. (D) Western blot of endogenous NSUN2 in HepG2 after transfection with pAHC-Core, pAHC-Pol, and pAHC-preC. The expression of HBV Pol and Core were analyzed with antibody against HA. The expression of HBeAg was analyzed with ELISA. (E) Western blot of endogenous NSUN2 in HepG2 after transfection with pAHC-Core and pAHC-Core-C2017A. (F) qPCR results of NSUN2 mRNA levels after core protein transfection in HepG2 or infection in PHHs. GAPDH was used as the internal reference. (G) NSUN2 expression level in a published dataset from healthy or HBV infected human liver tissue (GEO: GSE83148). Immunoblots shown are representative of three independent experiments. Graphs show the mean ± SD derived from three independent experiments and were analyzed by one-way ANOVA analysis or unpaired Student’s t test. ns, not significant for P > 0.05, *P < 0.05, ***P< 0.001, ****P< 0.0001.