FIG 2.

PDCoV nsp5 cleaves pDCP1A depending on its protease activity. (A) HEK-293T cells were cotransfected with expression constructs encoding wild-type PDCoV nsp5 or its protease-defective mutants (H41A or C144A) and pDCP1A expression plasmid. After 28 h, cells were lysed for Western blotting. (B) HEK-293T cells were cotransfected with pCAGGS-nsp5-HA and pCAGGS-Flag-pDCP1A. After 24 h, cells were treated with MG132 (final concentration of 20 μM), Z-VAD-FMK (final concentration of 20 μM), or 3-MA (final concentration of 5 mM) for another 8 h. Cells were then lysed for Western blotting. WT, wild type.