Figure 4. Bim deletion rescues the... | Rockefeller University Press

Figure 4.

$Figure 4. Bim deletion rescues the generation of IELps in RasGRP1-deficient mice. RasGRP1−/− mice were crossed onto the Bim−/− background to create DKO mice. (A) Thymi were harvested from the indicated mice and analyzed for the presence of IELp cells. Total thymocytes were analyzed for expression of CD4 and CD8 and CD4−/dullCD8−/dull were gated (left). NK1.1− cells were subsequently analyzed for expression of TCRβ and CD5 (right). (B) Number of IELp cells (CD4−/dullCD8−/dullNK1.1−TCRβ+CD5+) from the indicated mice. Data were obtained from the indicated number of mice over 17 individual experiments; WT (n = 17), 1KO (n = 11), BKO (n = 7), DKO (n = 6). (C) Representative expression of CD122 on IELp cells (CD4−/dullCD8−/dullNK1.1−TCRβ+CD5+; left) and representative PD-1 and Helios expression on CD122+ IELp cells (right) from the indicated mice. (D) Frequencies of PD-1+Helios+ cells within CD122− and CD122+ IELp fractions from the indicated mice. Data were obtained from the indicated number of mice over nine individual experiments; WT (n = 9), 1KO (n = 5), BKO (n = 5), DKO (n = 6). (E) Percentages of active caspase 3+ cells within the CD122− and CD122+ IELp fractions from the indicated mice. Data were obtained from the indicated number of mice over 12 individual experiments; WT (n = 11), 1KO (n = 8), BKO (n = 4), DKO (n = 6). *, P < 0.05; **, P < 0.01; ***, P < 0.001 (unpaired Student’s t test). Error bars indicate SEM.$

Bim deletion rescues the generation of IELps in RasGRP1-deficient mice. RasGRP1^−/− mice were crossed onto the Bim^−/− background to create DKO mice. (A) Thymi were harvested from the indicated mice and analyzed for the presence of IELp cells. Total thymocytes were analyzed for expression of CD4 and CD8 and CD4^−/dullCD8^−/dull were gated (left). NK1.1⁻ cells were subsequently analyzed for expression of TCRβ and CD5 (right). (B) Number of IELp cells (CD4^−/dullCD8^−/dullNK1.1⁻TCRβ⁺CD5⁺) from the indicated mice. Data were obtained from the indicated number of mice over 17 individual experiments; WT (n = 17), 1KO (n = 11), BKO (n = 7), DKO (n = 6). (C) Representative expression of CD122 on IELp cells (CD4^−/dullCD8^−/dullNK1.1⁻TCRβ⁺CD5⁺; left) and representative PD-1 and Helios expression on CD122⁺ IELp cells (right) from the indicated mice. (D) Frequencies of PD-1⁺Helios⁺ cells within CD122⁻ and CD122⁺ IELp fractions from the indicated mice. Data were obtained from the indicated number of mice over nine individual experiments; WT (n = 9), 1KO (n = 5), BKO (n = 5), DKO (n = 6). (E) Percentages of active caspase 3⁺ cells within the CD122⁻ and CD122⁺ IELp fractions from the indicated mice. Data were obtained from the indicated number of mice over 12 individual experiments; WT (n = 11), 1KO (n = 8), BKO (n = 4), DKO (n = 6). *, P < 0.05; **, P < 0.01; ***, P < 0.001 (unpaired Student’s t test). Error bars indicate SEM.