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. 2012 Feb 10;31(7):1679–1691. doi: 10.1038/emboj.2012.18

Figure 4.

Figure 4

Endogenous levels of cIAP1, and ectopic expression of cIAP2, protect MEFs from killing by TNF much more than XIAP. (A) Sensitivity of gene deleted lines to induction of cell death. Cells described in Figure 3A were incubated for 24 h with 100 ng/ml Fc-TNF or 20 ng Fc-CD95L, 500 nM SM, 1 μM etoposide or 5 μg/ml cisplatin where indicated. Cells were stained with PI and analysed by flow cytometry. The mean+s.e.m. of 3–11 independent experiments is shown. (B, C) Ability of ectopically expressed IAPs to protect cIap2−/−cIap1−/− DKO cells. cIap2−/−cIap1−/− MEFs were complemented with inducible mouse cIAP1, FLAG–cIAP2, or mouse XIAP and induced with 10 nM 4-HT for 24 h, and then incubated with or without 100 ng/ml Fc-TNF for a further 24 h. Cells were stained with PI and analysed by flow cytometry. The mean values+s.e.m. of 5–14 independent experiments are shown (* indicates a non-specific band). (D, E) Ability of ectopically expressed IAPs to protect Xiap−/−cIap1−/− DKO cells. Xiap−/−cIap1−/− MEFs were complemented with inducible mouse cIAP1 or EGFP–XIAP, induced with 10 nM 4-HT for 24 h and then treated with 100 ng/ml Fc-TNF or 20 ng/ml Fc-CD95L and 500 nM SM. The mean values+s.e.m. of three to six independent experiments are shown. See also Supplementary Figure S2. Figure source data can be found in Supplementary data.