Fig. 5. Association between BANK and Lyn. (A and B) HEK293T cells were transiently transfected with combinations of plasmids expressing various FLAG-tagged mutant BANK and Lyn mutants. Anti-FLAG (BANK) and anti-Lyn immunoprecipitates were subjected to immunoblotting with anti-pTyr, anti-BANK and anti-Lyn antibodies. (C) HEK293T cells were transiently transfected with plasmids expressing FLAG-tagged BANK, with (+) or without (–) expressing LynY508F. The lysates were incubated with 1 µg of the GST fusion proteins indicated immobilized on glutathione– Sepharose. The precipitates were washed and immunoblotted with anti-BANK antibody (a). The amount of BANK in HEK293T cell lysates (b). The amount and purity of GST fusion proteins were verified by Coomassie Blue staining (c; M, molecular weight standards). Abbreviations are explained in the text.