Fig. 1: Ponatinib is a RAF inhibitor.
a Schematic representation of ERK signaling pathway from different BRAF species. b Schematic representation of the in-cell-western screening assay. c Screening of a library of 200 kinase inhibitors using in-cell-western assay. SKMEL239-C4 melanoma cells left untreated (regular media), treated with 0.5 μM Vemurafenib, 0.1 μM Trametinib, or with 5 μM of known RAF, MEK, and other kinase inhibitors in the presence of 0.5 μM Vemurafenib for 3 h. d SKMEL239-C4 melanoma cells left untreated (regular media), treated with 0.5 μM Vemurafenib, 0.1 μM Trametinib, Ponatinib 5 μM without or with 0.5 μM Vemurafenib for 3 h, and assayed with in-cell-western. Data are mean of n = 2 independent experiments. e Chemical structure of Ponatinib. f BRAF kinase activity assay in the absence or presence of Ponatinib or Vemurafenib was assayed by western blot with the indicated antibodies. g CRAF kinase activity assay in the absence or presence of Ponatinib or Vemurafenib was assayed by western blot with the indicated antibodies. g Kinase activity inhibition profiles of BRAFV600E and BRAFWT upon Ponatinib titration using SelectScreen assay. Data are mean of two technical replicates from n = 2 independent experiments. Source data are provided as a Source Data file.
