TEL-JAK2 expression results in constitutive tyrosine phosphorylation of Shc and association with the SH2 domain of Shc.

(A) Ba/F3 and Ba/F3-TEL-JAK2 cell lines were depleted of cytokine and then stimulated in the presence (+) or absence (−) of IL-3. IPs were performed using a Shc antibody, and tyrosine-phosphorylated proteins were detected by pTyr IB (upper panel). Total Ship-1, Grb2, and Shc proteins were detected upon reprobing with Ship-1, Grb2, and Shc antibodies, respectively (lower panels). (B) Ba/F3 and Ba/F3-TEL-JAK2(5-19) cells were depleted of cytokine and then stimulated in the presence (+) or absence (−) of IL-3. GST in vitro mixes were performed using 5 μg GST (lanes 1-4), GST-Shc PTB domain (lanes 5-8), GST-Shc CH1 domain (lanes 9-12), or GST-Shc SH2 domain (lanes 13-16). Tyrosine-phosphorylated proteins were detected by pTyr IB.