FIG 5.

HDAC6-mediated degradation of PDCoV nsp8 is dependent on its deacetylase activity. (A and B) LLC-PK1 cells were cotransfected with pCAGGS-HA-nsp8 and pCAGGS-Flag-HDAC6 or empty vector, along with or without treatment with tubacin (5 μM) (A) or TSA (1 μM) (B) for 8 h. The cells were collected and subjected to Western blotting for detecting HA-nsp8, Flag-HDAC6, Ac-α-tubulin, α-tubulin, and β-actin. (C) Schematic diagram of porcine HDAC6 (pHDAC6) and pHDAC6 deacetylase dead mutants. (D) LLC-PK1 cells were cotransfected with pCAGGS-HA-nsp8 and expression plasmids encoding HDAC6-WT, HDAC6-DM, HDAC6-H258A, HDAC6-H652A, or empty vector for 28 h. The cell lysates were subjected to a Western blotting assay.