Figure 2.
The Jak2S523L mutation causes IL-3–independent growth in Ba/F3 cells expressing EPOR through Jak2/Stat5 activation by impairing phosphorylation of S523, a negative regulatory site for Jak2. (A) Ba/F3 cells transduced with MPL and either Jak2 WT, Jak2S523L, Jak2V617F, or Jak2K539L were grown in the absence of IL-3. Results of 3 independent experiments are depicted as means ± standard errors of the mean. (B) Ba/F3 cells transduced with EPOR and either Jak2 WT, Jak2S523L, Jak2V617F, or Jak2K539L were grown in the absence of IL-3. Results of 3 independent experiments are depicted as means ± standard deviations. (C) WB analysis of Jak2/Stat5, Akt and mitogen-activated protein kinase (Mapk) signaling in Ba/F3-MPL or Ba/F3-EPOR cells transduced with Jak2 WT, Jak2S523L, Jak2V617F, or Jak2K539L. (D-E) Lysates from Ba/F3 cells were immunoprecipitated (IP) with total Jak2 antibody and analyzed by WB using antibodies for pS523 and pY570, respectively.

The Jak2S523L mutation causes IL-3–independent growth in Ba/F3 cells expressing EPOR through Jak2/Stat5 activation by impairing phosphorylation of S523, a negative regulatory site for Jak2. (A) Ba/F3 cells transduced with MPL and either Jak2 WT, Jak2S523L, Jak2V617F, or Jak2K539L were grown in the absence of IL-3. Results of 3 independent experiments are depicted as means ± standard errors of the mean. (B) Ba/F3 cells transduced with EPOR and either Jak2 WT, Jak2S523L, Jak2V617F, or Jak2K539L were grown in the absence of IL-3. Results of 3 independent experiments are depicted as means ± standard deviations. (C) WB analysis of Jak2/Stat5, Akt and mitogen-activated protein kinase (Mapk) signaling in Ba/F3-MPL or Ba/F3-EPOR cells transduced with Jak2 WT, Jak2S523L, Jak2V617F, or Jak2K539L. (D-E) Lysates from Ba/F3 cells were immunoprecipitated (IP) with total Jak2 antibody and analyzed by WB using antibodies for pS523 and pY570, respectively.

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