Figure S2.
S161 phosphorylation is required for TNF-induced RIPK1 kinase–dependent apoptosis, but not RIPK1 kinase–independent apoptosis, in CT26 cells and in primary BMDMs. (A–D)Ripk1-deficient CT26 cells reconstituted with Flag-RIPK1 WT or mutants (A) and primary BMDMs from three mice of each genotype as indicated (B–D) were treated with TNF (10 ng/ml), TNF (10 ng/ml) + SMAC mimetic (1 μM), and TNF (10 ng/ml) + CHX (1 μg/ml), respectively, in the presence or absence of GSK-872 for the indicated time points. All mice were 8- to 12-wk-old male littermates. Cell survival was measured. Data are presented as mean ± SD. Cell lysates of nontreated cells were analyzed by immunoblotting to detect proteins as indicated. The above experiments were independently performed twice. Source data are available for this figure: SourceData FS2. Refer to the image caption for details.

S161 phosphorylation is required for TNF-induced RIPK1 kinase–dependent apoptosis, but not RIPK1 kinase–independent apoptosis, in CT26 cells and in primary BMDMs. (A–D) Ripk1-deficient CT26 cells reconstituted with Flag-RIPK1 WT or mutants (A) and primary BMDMs from three mice of each genotype as indicated (B–D) were treated with TNF (10 ng/ml), TNF (10 ng/ml) + SMAC mimetic (1 μM), and TNF (10 ng/ml) + CHX (1 μg/ml), respectively, in the presence or absence of GSK-872 for the indicated time points. All mice were 8- to 12-wk-old male littermates. Cell survival was measured. Data are presented as mean ± SD. Cell lysates of nontreated cells were analyzed by immunoblotting to detect proteins as indicated. The above experiments were independently performed twice. Source data are available for this figure: SourceData FS2.

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