Fig. 1: Identification of Vemurafenib as a new necroptosis inhibitor.
From: Vemurafenib inhibits necroptosis in normal and pathological conditions as a RIPK1 antagonist
A Scheme of the in vitro high-throughput drug screen workflow. L929 cells were treated with each compound (10 µM) and then stimulated with TNFα (20 ng/mL) for 24 hours. Cell viability was determined by CellTiter-Glo® assay. B Identification of necroptosis inhibitors by cellular screen with drug library. Normalized necroptosis inhibition efficiency for drugs tested depicted as a percentage of control (Nec-1 = 100). Data are represented as the mean of triplicates. The chemical structure of Vemurafenib is shown above. MDF (C) and HT29 (D) cells were pretreated with Promethazine hydrochloride (10 μM; PH), Lbrutinib (10 μM; Lbr) or Vemurafenib (10 μM; Vem) followed by treatment with TNFα (20 ng/mL), Smac mimetic (1 μM), and z-VAD (20 μM; TSZ) for 6 and 12 hours, respectively. Cell viability was determined by CellTiter-Glo® assay. E Representative images of L929, MDF, and HT29 cells treated with DMSO, Vemurafenib, or Nec-1 followed by necroptosis induction. Necroptosis in L929 cells was induced by TNFα for 24 hours. Necroptosis in MDF and HT29 cells was induced by TNFα, Smac mimetic, and z-VAD for 6 and 12 hours, respectively. F Dose-response curves showing the inhibitory effect of Vemurafenib on necroptosis in L929, MDF, and HT29 cells. MDF (G) and HT29 (H) cells were pretreated with DMSO, Vemurafenib (30 μM), or Nec-1 (10 μM) followed by indicated treatment for 6 and 12 hours, respectively. Cell viability was determined by CellTiter-Glo® assay. TS: TNFα (20 ng/mL) plus Smac mimetic (1 μM). P value was calculated by unpaired Student’s t-test. (*p < 0.05, **p < 0.01, ***p < 0.001).
