FIG 5.

SUMOylation regulates the stability of NS5. (A and B) BHK-21 cells were transfected with WT or SIM-mutated NS5 for 24 h and then treated with cycloheximide (CHX). Cells were harvested at different time points after CHX treatment as indicated. (A) The NS5 protein expression levels were analyzed by WB using antibodies against HA and actin. (B) The band intensity of NS5 was quantified by ImageJ and normalized to that of actin. The relative quantities (RQ) were calculated and are shown as percentages of the quantities at 0 h. (C) BHK-21 cells were transfected with SIM-mutated NS5 for 24 h, designated hour 0, and then either left untreated or treated with CHX (lanes 2 to 6) together with the indicated inhibitors for another 3 h. Cells were harvested and analyzed by WB with anti-HA and anti-actin antibodies.