FIGURE 2.

TRIM32 knockdown makes cells resistant to TNFα-induced apoptosis. HEK293T cells were infected with lentiviral constructs for expressing TRIM32 shRNA or control shRNA, and the infected cells were pooled after 1 μg/ml puromycin selection. The cells were then treated with 50 ng/ml TNFα and 100 ng/ml cycloheximide (CHX) for 12 h. A, cell lysates were analyzed by immunoblotting with an anti-TRIM32 antibody to detect TRIM32 expression levels and with an anti-cleaved caspase-3 antibody to detect caspase-3 activity. ns, nonspecific. B, caspase-3 activity was also detected with a fluorometric assay Data are expressed as the mean ± S.D. of three independent experiments. C, cell morphologies were observed under a microscope.