Fig. 3

RIPK1/RIPK3/MLKL necrosome is required for hyperphosphorylated tau-stimulated cytokine induction. A, D, G NC HT-22 and RIPK1-KO HT22 cells were transfected with vector or TauP301S. A Lysates were analyzed by western blotting with the indicated antibodies. D LDH was assayed to determine cell death. G Levels of indicated cytokines were analyzed by qPCR (B, E, H) NC HT-22 and RIPK3-KO HT22 cells were transfected with vector or TauP301S. B Lysates were analyzed by western blotting with indicated antibodies. E LDH was assayed to examine cell death. H Levels of indicated cytokines were analyzed by qPCR. C, F, I NC HT-22 and MLKL-KO HT22 cells were transfected with vector or TauP301S. C Lysates were analyzed by western blotting using the indicated antibodies. F LDH was assayed to examine cell death. I Levels of indicated cytokines were analyzed using qPCR Data are presented as the mean ± standard error of the mean (SEM) of three experiments, and a two-way ANOVA with Tukey’s multiple comparisons test was used to analyze the statistical significance of the data