Figure 6

Gab1 phosphorylation is required for PTP1B binding. Myc-tagged Gab1 was expressed into HEK-293 T cells followed by pervanadate treatment. The proteins were incubated either in the presence or absence of wild type GST-PTP1B followed by pull-down assay with either wild type PTP1B or mutant PTP1B. The bound proteins were subjected to immuno blot analysis with anti-Myc antibody (A). The blot was reprobed with anti-GST antibody to ensure equal amount of fusion in each pull-down (A). Myc-tagged Gab1 expressed in HEK-293 T were either treated or untreated with pervanadate. Pervanadate treated or untreated samples were subjected to GST pull-down assay with either wild type PTP1B or mutant PTP1B followed by immuno blot analysis with either Gab1 or pGab1 (B). The blot was reprobed with anti-GST antibody to ensure equal amount of fusion in each pull-down (B).