Analysis of Bim phosphorylation in JAK2
V617F mutant SET-2 cells. A: SET-2 cells were treated with DMSO or 500 nM NVP-BSK805 for 24 hours. Bim was immunoprecipitated and levels of Bim-EL Ser69 as well as Ser59 phosphorylation were analyzed by Western blotting. Levels of P-STAT5 and total STAT5 were probed for as controls for the drug treatment. B: SET-2 cells were treated with DMSO, 500 nM NVP-BSK805 or 10 μM UO126 for 4 hours. Levels of P-STAT5, STAT5, P-ERK1/2 and ERK1/2 were assessed by Western blotting. Bim was immunoprecipitated and levels of Bim-EL Ser69 phosphorylation were detected by Western blotting.