Critical role of RasGRP1 in TCR-induced mTORC1, mTOR2, and PI3K/Akt activation. Thymocytes from WT (Wt) and RasGRP1 KO (Grp1 KO) mice were left unstimulated or stimulated with anti-CD3 (500A2) for 5 minutes at 37°C. Cell lysates were separated by SDS-PAGE followed by immunoblotting with the indicated antibodies. The blots were stripped and reprobed with an anti–β-actin antibody for a loading control. Data shown and bar graphs are representative of/quantified from 3 experiments. *P < .05; **P < .01; ***P < .001 indicate statistical significance between WT and RasGRP1KO after anti-CD3 stimulation.