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. 2020 Aug 12;9(8):1889. doi: 10.3390/cells9081889

Figure 3.

Figure 3

HBV release requires COPII components. HuH-7 cells were treated with control siRNA (siCon) or siRNAs targeting Sec24A, Sec24B, Sec23A, Sec23B, Sar1A or Sar1B for 48 h and retransfected with the pHBV* replicon. After an additional 72 h, lysates and supernatants were harvested. To check for depletion, transcript levels of the RNA interference (RNAi)-targeted genes were measured by qRT-PCR and demonstrated in percent amount relative to control cells (n = 4, ± SD). To monitor protein synthesis, cell lysates were examined by WB using tubulin-, L- (K1350) and core-specific (K46) antibodies. The nonglycosylated (p39) and glycosylated (gp42) forms of L are indicated. HBV release was measured by envelope-specific IP of supernatants using anti-L (K1350) and anti-S (K38) antibodies and real-time multiplex PCR measurements of the viral genomes (extracellular virions). Nonenveloped intracellular nucleocapsids (intracellular NCs) were immunoprecipitated with anti-capsid (K45) antibodies and analyzed by PCR. PCR results were demonstrated in the percent amount relative to siCon-transfected cells. Error bars indicate the standard deviations from the mean of three experiments measured in duplicates. ** p < 0.01 compared to control.