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. 2004 May;24(10):4557–4570. doi: 10.1128/MCB.24.10.4557-4570.2004

FIG. 10.

FIG. 10.

Tyrosine 813 is required for myc-SH2-Bβ(504-670) to enhance JAK2-mediated phosphorylation of STAT5B. COS7 cells were cotransfected with 2.0 μg of cDNA encoding STAT5B and 2.0 μg of either empty vector (lane 1) or cDNA encoding myc SH2-Bβ(504-670) (lanes 2, 4, and 6), without (lanes 1 and 2) or with 0.25 μg of cDNA encodes either JAK2 (lanes 3 and 4) or JAK2 (Y813F) (lanes 5 and 6). Cell lysates were immunoblotted with anti-P-STAT5 (first panel), anti-STAT5B (second panel), and anti-myc (third panel). Cell lysates were also immunoprecipitated with anti-JAK2 and the precipitated proteins were immunoblotted with anti-JAK2 (fourth panel) and with anti-PY (fifth panel). The nitrocellulose membrane was stripped between blots as outlined in Materials and Methods.