Fig. 5.

Selective TAK1 loss augments the activation of RIPK1 to promote apoptosis. (A) WT, MAP3K7^fl/fl; Lyz2^cre/+, and MAP3K7^fl/fl; Emx^cre/+ mice (each dot representing 1 mouse) were subjected to transient 60 min of MCAO, followed by reperfusion for 23 h. The brains were processed for TTC staining to measure infarct volume. (B) Brain lysates from mice with the indicated genotypes subjected to MCAO for 60 min followed by reperfusion for the indicated time periods were immunoprecipitated with p-S166 RIPK1 and analyzed by immunoblotting with the indicated antibody. (C–E, Upper) Sections of WT, MAP3K7^fl/fl; Lyz2^cre/+, and MAP3K7^fl/fl; Emx^cre/+ mice treated with transient 60 min of MCAO followed by reperfusion for 5 h (C and E) or 3 h (D) at different time points as indicated on the bar graphs were immunostained with p-S166 RIPK1 and coimmunostained with IBA1, CD31, or NeuN for microglia/infiltrated macrophages (C), cerebrovascular endothelial cells (D), and neurons (E), respectively. The larger Insets (Right) in C are an enlarged image of the smaller Insets (Left). (Magnification: 40×; Scale bar: 50 μm.) (C–E, Lower) The quantification of the results. The data represent mean ± SEM. The quantifications of p-S166 Ripk1⁺ microglia/infiltrated macrophages, endothelial cells, and neurons are shown at the bottom of each panel. n = 3 mice for each time point. Assessment of each time point includes data from five sections for approximately 150 cells in each mouse. *P < 0.05 and **P < 0.01.