Figure 6.
EBOV-GP is ubiquitinated and degraded in the presence of PDIA3. (a) GP was expressed with WT Ub or indicated Ub mutants in HEK293T cells in the presence or absence of PDIA3. Protein expression was analyzed by WB. (b) GP was expressed with WT Ub or indicated Ub mutants in HEK293T WT and PDIA3-KO cells. Protein expression was analyzed by WB. (c) GP was expressed with increasing amounts of WT Ub or Ub[7 K/R] in the presence or absence of PDIA3 in HEK293T cells. Protein expression was analyzed by WB. (d) GP was expressed with UbK48,63R in the presence or absence of PDIA3 in HEK293T cells. GP was pulled down by anti-FLAG and protein expression in cell lysate (Input) and IP samples was analyzed by WB. (e) GP was expressed in the presence or absence of PDIA3 in HEK293T cells. Cells were treated with 50 μM cycloheximide (CHX) in the presence or absence of 100 nM BafA1. Cells were collected at indicated time points and protein expression was analyzed by WB. The levels of GP in A, B, C, D, and E were further quantified. Error bars represent SEMs calculated from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns (p > 0.05).