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. 2021 Nov 8;36(3):701–711. doi: 10.1038/s41375-021-01462-4

Fig. 5. Phosphorylation of CSF2RB by FLT3-ITD requires only one phosphorylated tyrosine within the JMD and takes place regardless of ITD sequence and insertion site.

Fig. 5

A Ba/F3 cells were transduced with CSF2RB-Flag and either FLT3 or FLT3-ITD 598/599 [12] containing mutations of tyrosines 589 and 591 to phenylalanine as indicated. Cells were serum-deprived for 5 h and co-immunoprecipitations were performed using anti-flag beads capturing CSF2RB. Immunoprecipitates and whole-cell lysates were subjected to SDS–PAGE and western blot analysis using indicated antibodies. B Parental Ba/F3 cells and Ba/F3 cells expressing CSF2RB-Flag in combination with either FLT3 or one FLT3-ITD variant were serum-deprived for 5 h. Co-immunoprecipitations were performed using anti-flag beads capturing CSF2RB. Immunoprecipitates and whole-cell lysates were subjected to SDS–PAGE and western blot analysis using indicated antibodies. Sequences and insertion sites of utilized FLT3-ITD variants are depicted on the right.