Cbl Enforces Vav1 Dependence and a Restricted Pathway of T Cell Development
Figure 3
Phosphorylation of normally Vav1-dependent signaling molecules is enhanced in Cbl-/-Vav1-/- thymocytes.
DP thymocytes were sorted from WT, Cbl-/-, Vav1-/- and Cbl-/-Vav1-/- mice by flow cytometry, cultured without stimulation (Ctrl) or with anit-CD3 Ab (Stim) for 2 minutes at 37°C. and protein lysates prepared. A) Protein lysates were immunoprecipitated (IP) with anti-PLC γ1 and immunoblotted (IB) with anti-phosphotyrosine (4G10) (upper panel) or anti-PLC γ1 (lower panels). The results presented are representative of 4 experiments. B) Analysis as in Panel A, but with samples run and exposed to allow visualization and comparison of WT and Vav-/- DP thymocytes. C) Protein lysates were immunoblotted (IB) with anti-phospho-ERKs (upper panel) or anti-ERK (lower panel). The results presented are representative of 4 experiments.