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. 2022 Jul 16;13(7):618. doi: 10.1038/s41419-022-05076-1

Fig. 5. TAB2 is required for the resistance to TNF-induced necroptotic cell death in PPP6C deficient cells.

Fig. 5

A, B Control and Tab2-knockdown L929 cells were treated with TZ for the indicated time, and analyzed under a microscope (A) or quantified by flow cytometer (B) after stained with PI. Scale bar, 50 μm. C Control and Tab2-knockdown L929 cells were treated with TZ for the indicated time and then lysed for immunoblot with indicated antibodies. D, E Control and Tab2-knockdown Ppp6c-KO L929 cells were treated with TZ for the indicated time, and analyzed under a microscope (D) or quantified by flow cytometer (E) after stained with PI. Scale bar, 50 μm. F Control and Tab2-knockdown Ppp6c-KO L929 cells were treated with TZ for the indicated time and then lysed for immunoblot with indicated antibodies. G, H Control and Tab2-knockdown L929 cells were pretreated with TAK1 inhibitor (5Z-7-Oxozeaenol, 1 μM) in the presence or absence of Necrostatin-1 (10 μM) for 30 min, and then treated with TZ for 4 h. Cells were stained with PI and analyzed under a microscope (G) or quantified by flow cytometer (H). Scale bar, 50 μm. I Control and Tab2-knockdown L929 cells were pretreated with TAK1 inhibitor (5Z-7-Oxozeaenol, 1 μM) for 30 min, and then treated with TNF (10 ng/ml) for the indicated time. Cell lysates were probed with indicated antibodies. Data shown are representative of three independent experiments and presented as means ± SDs of triplicates (B, E, H). **p < 0.01, ***p < 0.001, with one-way ANOVA analysis (B, E, H).