Fig. 5: Quizartinib inhibited necroptosis through indirectly regulating RIPK1 kinase activity.
From: Repurposing of Ibrutinib and Quizartinib as potent inhibitors of necroptosis
a HT-29 cells were pretreated with vehicle, GSK’872 (10 μM), Quizartinib (10 μM) for 1 h following treatment with TSZ. Then the cell viabilities were determined using CCK8 method. n = 3 biologically independent samples. b HT-29 cells were pretreated with vehicle or Quizartinib (10 μM) for 1 h following treatment with TSZ for the indicated times. Then the cells were harvested and analyzed with the indicated antibodies. c Mouse RIPK3-flag reconstituted RIPK3-KO L929 cells were pretreated Quizartinib (2 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. d L929 cells were pretreated Quizartinib (2 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. e L929 cells were pretreated vehicle or indicated compounds for 1 h following treatment with TSZ. Then the cell viabilities were determined using CCK8 method. n = 3 biologically independent samples. f Plasmid expressing HA-RIPK1 or Vector was transfected into 293T cells. Then the cells were treatment with indicated concentrations of Quizartinib, NEC-1 (10 μM) or vehicle for 20 h. The cells were harvested and analyzed with indicated antibodies. Data shown are representative of three independent experiments. Means ± SDs. #p < 0.01.
