Figure 8.

Degradation of PCM1 by ZIKV in SK-N-SH cells. A, Western blotting assay to examine the levels of PCM1 and Mib1 in SK-N-SH cells that are infected or mock-infected with ZIKV at an MOI of 1 in the presence or absence of MG132 (10 μm). The MG132 was added 12 h before sample collection. NS3 shows viral infection, and tubulin is for control of sample loading. B, Western blotting assay to determine the dose dependence of MG132 in inhibiting the degradation of PCM1 by ZIKV. SK-N-SH cells were infected with ZIKV at an MOI of 1 for 24 h. The MG132 was added 12 h before collecting the samples. C, immunoprecipitation and Western blotting assays to determine ubiquitination of PCM1. The whole-cell lysate from mock-infected SK-N-SH cells, ZIKV-infected SK-N-SH cells, or ZIKV-infected SK-N-SH cells treated with TAK-243 (MLN7243) (purchased from Chemietek, catalog no. AOB87172) was incubated with anti-PCM1 and protein G beads; the beads were washed; and the eluted samples were probed with anti-PCM1 or anti-ubiquitin antibody.