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Figure 5


Fig. 5. FasL is phosphorylated by Fgr binding to the PRD. (A) Western blots of lysates from stably transfected RBL clones expressing equivalent levels of FasL-GFP (WT) and the GFP-tagged FasL mutants (KKR, {Delta}-Pro and 3Y), immunoprecipitated (IP) with anti-FasL antibody MIKE 1 and Protein-G-Sepharose or Protein-G-Sepharose alone (-). The blot was probed with anti-phosphotyrosine ({alpha}PY) antibody 4G10 (upper panel) and stripped and re-probed with anti-FasL antibody G247-4 (lower panel). Arrows denote the positions of bands corresponding to phosphorylated FasL-GFP (P-FasL-GFP), FasL-GFP and the {Delta}-Pro mutant as well as IgG heavy chain (IgGHc). (B) Immunotyrosine blot of HeLa cells expressing untagged WT and {Delta}-Pro FasL and Fgr-YFP (+) as indicated probed with anti-phosphotyrosine ({alpha}PY) antibody 4G10 (upper panel) and stripped and re-probed with anti-FasL antibody G247-4 (lower panel). Arrows denote the positions of bands corresponding to phosphorylated Fgr (P-Fgr-YFP), FasL-GFP (P-FasL-GFP), FasL-GFP and the {Delta}-Pro mutant. Positions of molecular masses are shown (kDa).