FIGURE 7.
Overexpression of p22-FLIP and cFLIPS results in RIP1 ubiquitination and its association with IKKγ. HEK293 cells were transduced with cFLIP variant or vFLIP lentivector. After 48 h, cells were lysed in a buffer containing 10 mm N-ethylmaleimide and 20 mm iodoacetamide to inhibit deubiquitinases. As a positive control, cells were treated with TNF-α (250 ng/ml) for 5 min. The cell lysates were immunoprecipitated (IP) with 2 μg of an anti-IKKγ (Ι) or an isotype-matched control antibody (C). The immunoprecipitates were analyzed by immunoblotting using the antibodies shown. Results are representative of two independent experiments. Ub, ubiquitin.