RasGRP1 KO and RasGRP1/3 DKO thymocytes display impaired proliferation of DN3 and inefficient transition from DN3E to DN3L. A.

<p>Intracellular TCRβ (TCRβ_i.c.) by forward scatter (FSC) profiles of DN3 (CD4⁻CD8⁻Thy1.2⁺CD44⁻CD25⁺) from B6 (n = 8), 1KO (n = 6), 3KO (n = 9) and DKO (n = 12) thymi. <b>B.</b> Frequencies of DN3 (CD4⁻CD8⁻Thy1.2⁺CD44⁻CD25⁺) and DN4 (CD4⁻CD8⁻Thy1.2⁺CD44⁻CD25⁻) expressing intracellular TCRβ (TCRβ_i.c.). <b>C.</b> Ratio of frequencies of TCRβ_i.c.⁺ DN3E/DN3L ((DN3E/DN3L)TCRβ_i.c.⁺). <b>D.</b> CD98 by CD25 profiles of Thy1.2⁺CD44⁻ cells from 1 and 2 day DN3E-OP9-DL1 co-cultures; data are representative of 3 independent experiments. <b>E.</b> Frequencies of BrdU⁺ DN3 (CD4⁻CD8⁻Thy1.2⁺CD44⁻CD25⁺) and DN4 (CD4⁻CD8⁻Thy1.2⁺CD44^–CD25⁻) from B6 (n = 5), 1KO (n = 5), 3KO (n = 6) and DKO (n = 7) mice injected with BrdU i.p. 2h prior to euthanasia. *p<0.05, **p<0.01 and ***p<0.001.