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

Fig. 2. Phosphatase activity of PPP6C is required for its role in regulating necroptosis.

Fig. 2

A, C Control and PPP6C-overexpression L929 cells were treated with TZ for 4 h, then stained with PI and analyzed under a microscope (A), or quantified by flow cytometer (C). Scale bar, 50 μm. B Control and PPP6C-overexpression L929 cells were treated with TZ for 4 h and cell lysates were probed with indicated antibodies. D, E Ppp6c-KO cells were transduced with PPP6C or its phosphatase dead (PD) lentivirus and followed by the treatment with TZ for the indicated time. Cells were then stained with PI and analyzed under a microscope (D) or quantified by flow cytometer (E). Scale bar, 50 μm. F, G Ppp6c-KO L929 cells were transduced with PPP6C lentivirus and followed by the treatment with TZ for the indicated time. Cell lysates were resolved on SDS-PAGE (F) or non-reducing PAGE (G) for immunoblot with indicated antibodies. H, I Ppp6c-KO L929 cells were transduced with PPP6C or PPP6C-PD lentivirus and followed by the treatment with TZ for the indicated time. Cell lysates were resolved on non-reducing PAGE (H) or SDS-PAGE (I) for immunoblot with indicated antibodies. Data shown are representative of three independent experiments and presented as means ± SDs of triplicates (C, E). **p < 0.01, ***p < 0.001, with an unpaired Student’s t-test (C) or one-way ANOVA analysis (E).