FIG 4.

HDAC6-mediated degradation of nsp8 is dependent on the ubiquitin proteasome pathway. (A) LLC-PK1 cells were cotransfected with fixed amounts of pCAGGS-HA-nsp8 and increasing quantities of pCAGGS-Flag-HDAC6 for 28 h, and then the cell lysates were analyzed by Western blotting with the indicated antibodies. (B to D) HEK-293T cells were cotransfected with pCAGGS-HA-nsp8 and pCAGGS-Flag-HDAC6 or empty vector and then treated by MG132 (20 μM) (B), Z-VAD (20 μM) (C), or 3-MA (5 mM) (D) for 8 h before collecting cell samples were collected and Western blotting assay was performed.