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. 2006 Sep 18;26(23):9035–9044. doi: 10.1128/MCB.00988-06

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Copyright © 2006, American Society for Microbiology

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FIG. 4. — Y239/Y240 tyrosines of ShcA regulate DN thymocyte development. (A) Analysis of thymic subsets based on CD4 versus CD8 (upper panels) or CD44 versus CD25 (lower panels) in ShcWT, ShcF317, ShcFF239/240, and ShcFFF transgenic mice in the lck-Cre background. Total thymocyte numbers are indicated on top. (B) STOP cassette deletion in thymocyte subsets and splenocytes analyzed by Southern blotting. The transgenic construct is revealed as a 7.1-kb or a 7.4-kb band (lane 1), while deletion of the STOP cassette by lck-Cre/Shc shifts the mobility to a 5.8-kb (ShcWT or ShcF317) or a 6.1-kb (ShcFFF) band. Lanes 1 and 14 show the nondeleted transgenes derived from mice not carrying Cre. These controls were run on the same gel but are placed adjacently here. (C) Lysates of total thymocytes or splenocytes from different ShcA transgenic mice (in the lck-Cre background) or a control (ShcFFF mouse without lck-Cre) were immunoprecipitated with anti-Flag monoclonal antibody and immunoblotted with anti-Shc antibody. Membranes were immunoblotted for actin to compare protein loading in the different lanes.