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Fig. 1. Abi1 is a downstream substrate of Bcr-Abl tyrosine kinase. (A) Ba/F3 and 32D cells were transduced with empty retroviral vector (control) or retroviral vector expressing p185Bcr-Abl (p185wt). Total lysates from 1x106 cells were analyzed by western blot (WB) using indicated antibodies. (B) Tyrosine phosphorylation of Abi1 in Bcr-Abl-transformed hematopoietic cells. Ba/F3 cells and the Ba/F3 expressing either p185wt or p185K671R were treated with or without 1 µM imatinib mesylate, as indicated. The lysates from 2x107 cells were immunoprecipitated (IP) with anti-Abi1 antibody, followed by western blot analysis using anti-phosphotyrosine (
-pTyr) antibody (upper panel). The blots were then stripped and reprobed with anti-Abi1 antibody (lower panel). The relative molecular mass (Mr) is indicated and presented as kilodaltons (kd).