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. Author manuscript; available in PMC: 2013 Apr 1.
Published in final edited form as: J Immunol. 2012 Mar 5;188(7):3031–3041. doi: 10.4049/jimmunol.1103226

FIGURE 2.

FIGURE 2

Requirements for Ly108 phosphorylation in the thymus: Contact mediated phosphorylation of Ly108. A. Schematic of lysis procedures. B. Ly108 was immunoprecipitated from (lane 1) intact thymi from C57Bl/6 mice directly lysed; (lane 2) thymocyte suspensions or (lane 3) thymocyte suspensions treated with pervanadate. Lysates were immunoprecipitated with Ly108 monoclonal antibody (top) or αTCR zeta (bottom) and blotted for phosphotyrosine (4G10), Ly108 and SAP (top) or phosphotyrosine and TCR zeta (bottom). C. Ly108 phosphorylation is reduced in absence of Fyn or SAP. Intact thymi from WT, Ly108−/−, Sh2d1a−/−, Fyn−/− and MHC Class I and II deficient mice were lysed, Ly108 immunoprecipitated and immunoblotted for 4G10, Ly108 and SAP. Relative intensities of pLy108/Ly108 and associated SAP/Ly108 are graphed. B and C are representative of 5 or more experiments.