FIG. 6.

ASFV DP71L(s) inhibits AFT4 and CHOP induction. (A) PK15 (lanes 1 to 3) and PK-DP71L(s)-V5 (lanes 4 to 6) cells were mock treated (lanes 1 and 4) or were treated with DTT for 30 min (lanes 2 and 5) or tunicamycin at 10 μg/ml for 4 h (lanes 3 and 6). Western blotting was used to analyze the cell lysates for the levels of total and phosphorylated eIF2α, ATF4, and CHOP proteins (indicated at the right). The total eIF2α was also used as the loading control. The lane numbers are indicated at the bottom. (B) Confocal microscopy was used to analyze the induction of CHOP in PK15 (top row) and PK-DP71(s)-V5 (bottom row) cells. The cells were treated with tunicamycin at 10 μg/ml for 4 h, and then they were permeabilized and costained with antibodies against V5 tag and CHOP, respectively, followed by appropriate Alexa Fluor 568- or 488-conjugated secondary antibodies. CHOP was stained green, and the V5 tag was stained red. The nucleus was stained blue with DAPI.