Fig. 3: Ibrutinib inhibited TNF-induced phosphorylation of RIPK3 in different cell lines.
From: Repurposing of Ibrutinib and Quizartinib as potent inhibitors of necroptosis
a HT-29, Hela-RIPK3, and NIH3T3-RIPK3 cells were pretreated with NEC-1 (20 μM), GSK’872 (10 μM), Ibrutinib (15 μM) or vehicle for 1 h following treatment with TSZ. Cell viabilities were determined using CCK8 method. For HT-29 and Hela-RIPK3 cells, n = 3 biologically independent samples. For NIH3T3-RIPK3 cells, n = 4 biologically independent samples. b, c HT-29 cells (b) or Hela-RIPK3 cells (c) were pretreated Ibrutinib (15 μM) or vehicle for 1 h following treatment with TSZ for indicated time. Then cells were harvested and analyzed with the indicated antibodies. d Mouse RIPK3-flag reconstituted RIPK3-KO L929 cells were pretreated Ibrutinib (15 μM) or vehicle for 1 h following treatment with TSZ for indicated time. Then cells were harvested and immunoprecipitated with M2 (anti-flag) antibody. The total cell lysates (TCL) and the immunoprecipitates were immunoblotted with the indicated antibodies. e L929 cells were pretreated Ibrutinib (15 μM) or vehicle for 1 h following treatment with TSZ for indicated time. Then the cells were lysed with Triton X-100 lysis buffer. The insoluble fractions were collected and analyzed with the indicated antibodies. Data shown are representative of three independent experiments. Means ± SD. *p < 0.05, #p < 0.01.
