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. Author manuscript; available in PMC: 2008 May 20.
Published in final edited form as: Exp Hematol. 2003 Dec;31(12):1310–1316. doi: 10.1016/j.exphem.2003.08.009

Figure 4.

Figure 4

Expression of Stat5b-DN abrogates EPO-dependent Stat5 activation by WT and mutant EPOR isoforms. (A): Whole-cell lysates from parental 32D cells (32D) or 32D cells coexpressing WT, 1–321, or YF EPORs and Stat5b-DN were analyzed by immunoblotting with an antibody specific for residues 451–649 of murine Stat5. The migration positions of endogenous Stat5 (Stat5) and Stat5b-DN (Stat5b-DN) are indicated. (B): 32D cells expressing WT, 1–321, or YF EPORs and lacking Stat5b-DN expression, and cell lines expressing WT, 1–321, or YF EPORs and Stat5b-DN were starved of exogenous growth factors in serum-free medium. Cells were left unstimulated or were treated with EPO (50 U/mL) for 10 minutes at room temperature. Stat5 was immunoprecipitated from detergent cell extracts and analyzed by immunoblotting with antiphosphotyrosine antibody 4G10 (upper panel). Blots were stripped and reprobed with Stat5-specific antibody (lower panel).