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. 2016 Apr 29;90(10):4889–4904. doi: 10.1128/JVI.00017-16

FIG 6.

FIG 6

Cellular STAT1 is not ubiquitinated by the presence of BoHV-1 VP8. (A to D) HEK293T cells were transfected with pFLAG/pHis-Ub, pFLAG-VP8/pHis-Ub, and pSV5V/pHis-Ub in the presence or absence of 10 μM MG132. At 24 h posttransfection, cell lysates were generated and incubated with His-60 nickel resin in the presence (A) or absence (C) of MG132 followed by Western blotting with anti-STAT1 antibody. Input lysates of pFLAG/pHis-Ub-, pFLAG-VP8/pHis-Ub-, and pSV5V/pHis-Ub-transfected cells are shown in panels B and D. (E and F) HEK293T cells were transfected with pFLAG or pFLAG-VP8. At 24 h posttransfection, cell lysates were generated and incubated with antiubiquitin antibody (E) and anti-VP8 antibody (F), followed by protein G-Sepharose. Monoubiquitinated VP8 was detected by Western blotting with anti-FLAG antibody (E) or with anti-Ub antibody (F).