Anti cleaved caspase 8

Western blotting analysis was performed as previously described69 (link). Briefly, tumor cells were cultured in the presence of the indicated ceramide analogs or ceramide analogs plus MegaFasL for 4 h. Cells were collected and lysed in cytosol buffer [10 mM Hepes, pH 7.4, 250 mM Sucrose, 70 mM KCl, 1.5 mM MgCl2, 1 mM EDTA, 1 mM EGTA, protease and phosphatase inhibitor cocktails (Calbiochem, Billerica, MA), and 0.01% digitonin] for 10 min. Cytosolic fractions were resolved in 4–20% SDS polyacrylamide gel and analyzed by Western blotting. Anti-cleaved caspase 8 and anti-cIAP1 were obtained from R&D Systems. Anti-cleaved human PARP, anti-xIAP, anti-p38, anti-p-p38, anti-pErk1/2, anti-Erk1/2, anti-pJNK, and anti-JNK antibodies obtained from Cell Signaling. β-actin was obtained from Sigma-Aldrich.

Western blotting analysis was performed as previously described [65 (link)]. Anti-cleaved caspase 8 (Cat#: AF705, 0.5μg/ml) and anti-cIAP1 (Cat#: AF8181, 1:500) were obtained from R&D systems. Anti-caspase 3 (Cat#: 9661, 1:1000), anti-caspase 9 (Cat#: 9501, 1:500), anti-PARP (Cat#: 9541, 1:250), anti-xIAP (Cat#: 2042, 1:500), anti-p100/p52 (Cat#: 4882, 1:1000), anti-IκBα (Cat#: 4814, 1:2000), anti-pIκBα (Cat#: 2859, 1:1000) antibodies were obtained from Cell Signaling. Anti-cIAP2 (Cat#: NB110-57030, 1:250) was obtained from Novus. Anti-β-actin (Cat#: A5441, 1:5000) was obtained from Sigma (St Louis, MO).