Figure 5: Association of ITK-Y511F and ITK-BTK(SH3) with SLP-76. (a) Jurkat cells that had been transfected with the indicated GFP-ITK constructs were stimulated (+) or not (−) with anti-CD3ε antibodies and then lysed. ITK in the lysates was immunoprecipitated (IP) with anti-GFP antibodies, immune complexes resolved by SDS-PAGE, proteins transferred onto PVDF membranes, and immunoblotted (IB) sequentially with anti SLP-76 (top panel) and anti-ITK (bottom panel) antibodies. Bands were visualized by chemiluminescence as described in Section 2. Numbers under each blot represent the association index between each ITK-variant and SLP-76 calculated as described in Section 2. Results are those from one of three replicate experiments. (b) average (±SEM) ITK-SLP-76 association index of three replicate experiments (including the experiment depicted in panel (a) performed as described in panel (a). Numbers in brackets denote values calculated by the Student’s t test.