PMC:7464876 / 58049-59812 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/7464876","sourcedb":"PMC","sourceid":"7464876","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7464876","text":"HBV L recruits envelopment-competent core/capsids to ER-associated structures. (A) HuH-7 cells were transfected with the wild-type (wt) HBV replicon construct (HBV) or mutant replicons defective in core protein expression (HBV.Cminus), envelope protein expression (HBV.Envminus), L envelope protein expression (HBV.Lminus) or polymerase activity (HBV.Polminus). Cells shown in panel F were cotransfected with the core-defective replicon and a plasmid encoding the core mutant C.K96A. PFA-fixed cells were double-stained with mouse antibodies against L (57761, L, green) and rabbit antibodies against core (B0586, C, red), followed by staining with Alexa Fluor 488-conjugated anti-mouse and Alexa Fluor 546-conjugated anti-rabbit antibodies (A–G). In the panels shown in E, cells were reacted with rabbit anti-L antibodies (K1350, L, green) and mouse antibodies recognizing the viral capsid rather than the core protein (3HB17, Capsid, red). Overlaid images are displayed in the Merge panels, and nuclei staining is shown in blue. Bar, 10 µM. Outlined areas are shown at larger magnifications. For quantitative colocalization analysis (QCA), Pearson´s correlation coefficient (PCC) values were calculated and represent the mean ± SD. (H) HuH-7 cells were transfected with HBV.Cminus together with control plasmids (Con), the wt core (C) or mutant C.K96A genes. Cell extracts were prepared by CHAPS-lysis and assayed by L- and core-specific WB to monitor protein expression (Input). For immunoprecipitation (IP), lysates were incubated with anti-core antibodies (K45), followed by immunoblotting with the L-specific MA18/7 antibody. IgG.HC denotes IgG heavy chains. Coimmunoprecipitation (CO-IP) analyses were done in triplicate, and a representative blot is shown.","tracks":[]}