Fig. 3: Phospho-specific antibody fail to immunoprecipitate JAK2 in the presence of ATP-competitive inhibitors. | Leukemia

Fig. 3: Phospho-specific antibody fail to immunoprecipitate JAK2 in the presence of ATP-competitive inhibitors.

From: Ruxolitinib mediated paradoxical JAK2 hyperphosphorylation is due to the protection of activation loop tyrosines from phosphatases

Fig. 3

JAK2-V617F expressing Ba/F3 cells were treated without and with vanadate, ruxolitinib and fedratinib. Cells were lysed in a non-denaturing lysis buffer, and immunoprecipitation was carried out using the anti-flag antibody, anti-JAK2 antibody and phosphospecific Tyr1007/Tyr1008 JAK2 antibody. Immunocomplexes and whole-cell lysate (WCL) were analyzed by IB (A). Denaturing immunoprecipitation was carried out using the same antibodies mentioned in non-denaturing immunoprecipitation, and these results showed all the antibodies were able to immunoprecipitate JAK2 equally either in the presence or in the absence of ATP-competitive inhibitors (B). A representative image of n = 2 two independent experiments is shown.

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