Fig. 6.

Transgenic T cells exhibit higher levels of inactive Fyn and Lck in LRs and fail to be activated after CD3/CD28 costimulation. a Western blotting and TLC of raft (R) and nonraft (NR) fractions prepared at rest (0 min) and after the activation (30 min) of transgenic and WT mouse T cells. Rafts are enriched in Flotillin-1 and cholesterol. Samples were analyzed with anti-pY⁵²⁸Fyn and anti-pY⁵⁰⁵Lck antibodies, which recognize the inactive forms of Fyn and Lck, respectively, followed by antibodies recognizing total Fyn and Lck, respectively. Cholesterol was analyzed by TLC. b, c Inactive/total Fyn and inactive/total Lck in raft microdomains according to densitometric analysis of the bands shown in a