Fig. 5

Functional enrichment-based clustering analysis for the quantified ubiquitylome. a, b L/H ratios for selected regulated Kub-sites & proteins from three biological replicates in HepG2.2.15 cells. c The levels of the secreted HBeAg and HBsAg were determined by ELISA from the cell culture supernatants samples co-transfected (1:1) with pHBV1.3 and flag-tagged TOLLIP. Co-immunoprecipitation and immunoblot analysis of extracts of Huh7 cells transfected with FLAG-tagged TOLLIP and HA-tagged wild type ubiquitin, Hepatitis B virus (HBV) 1.3-fold genome plasmid (pHBV1.3) and treated with MG132, are shown. d, e Co-immunoprecipitation and immunoblot analysis of Huh7 cells transfected with FLAG-tagged indicated plasmids together with HA-tagged wild type ubiquitin, Hepatitis B virus (HBV) 1.3-fold genome plasmid (pHBV1.3) and treated with MG132, are shown. f The levels of the secreted HBeAg and HBsAg were determined by ELISA from the cell culture supernatants samples co-transfected (1:1) with pHBV1.3 and flag-taged DNAJB6. Co-immunoprecipitation and immunoblot analysis of extracts of Huh7 cells transfected with FLAG-tagged DNAJB6 and HA-tagged wild type ubiquitin, Hepatitis B virus (HBV) 1.3-fold genome plasmid (pHBV1.3) and treated with MG132, are shown. g, h L/H ratios for selected regulated Kub-sites & proteins from three biological replicates in HepG2.2.15 cells. i The levels of the secreted HBeAg and HBsAg were determined by ELISA from the cell culture supernatants samples co-transfected (1:1) with pHBV1.3 and indicated plasmids. The data represent the average of three independent experiments and were analyzed with a two-tailed unpaired t test. Graphs indicate mean ± S.D. (n = 3) derived from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001