Fig. 5.

Transgenic T cells exhibit a hypophosphorylated protein profile with the downregulation of active Src. a Representative Western blot of protein lysates from transgenic and WT T cells after 60 min of activation by anti-CD3/CD28 antibodies incubated with anti-phosphotyrosine 4G10; blots were stripped and reprobed with anti-GAPDH antibody. b–e Western blots of protein lysates from transgenic and WT T cells several time points following anti-CD3/CD28 antibody activation (1 µg/ml each); blots were stripped and reprobed with an antibody raised against total specific protein. The results are representative of three independent experiments [pY⁴¹⁸Src/total Src: one-way ANOVA, *p = 0.0163; pY⁵⁰⁵Lck/total Lck: Kruskal-Wallis test, *p = 0.0362; pY⁵²⁸Fyn/total GAPDH, one-way ANOVA, *p = 0.0204, Tg vs. WT (30 min), **p = 0.0015; pYr³¹⁹Zap70/total Zap70, one-way Anova, **p = 0.0068]. f Immunofluorescence staining for pY⁴¹⁸Src in transgenic and WT T cells isolated by negative immunoselection