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. 2013 Nov 19;17(11):1397–1409. doi: 10.1111/jcmm.12156

Figure 4.

Figure 4

Effect of the JAK2/JAK1 inhibitor ruxolitinib and PI3K inhibitor NVP-BEZ236 alone or in combination on signalling in Ba/F3 TpoR cells overexpressing JAK2 wild-type (WT) or JAK2 V617F. Cells expressing TpoR JAK2 V617F were autonomous for growth, while Ba/F3 TpoR JAK2 WT cells were grown in medium supplemented with IL3. Cells were treated for the indicated intervals with 1 μM ruxolitinib and 10 μM NVP-BEZ235 alone or in combination and western blotting was performed with the phospho-specific antibodies (phospho-JAK2 (Tyr1007/1008), phospho-STAT3 (Tyr705), phospho-STAT5 (Tyr694), phospho-p70 S6 kinase (Thr389), phospho-S6 ribosomal protein (Ser235/236), phospho-AKT (Ser473), phospho-p38 MAP-kinase (Thr180/Tyr182), phospho-p44/42 ERK1/2 (Thr202/Tyr204), with antibodies to actin (as loading control) and with antibodies detecting the signalling proteins themselves.