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Autophosphorylation of Y980 is necessary for STAT5A phosphorylation. COS-7 cells were cotransfected with STAT5A cDNA (5 μg) and the cDNAs (5 μg) encoding: WT JAK3 (lane 1), K855A (lane 2), Y980F (lane 3), and Y981F (lane 4), or transfected with 5 μg STAT5A cDNA only (lane 5). Two days later, cells were lysed, immunoprecipitated with anti-STAT5A antiserum, and immunoblotted with mAb 4G10 (Upper) and anti-STAT5A antiserum (Lower).
