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. 2015 Jun 4;11(7):1052–1062. doi: 10.1080/15548627.2015.1055439

Figure 3.

Figure 3.

Inhibition of MTOR restored autophagy and survival in Tnfaip3-deficient T cells in vitro. (A) The effect of Torin1 on LC3 puncta formation in CD4 T cells. Naïve CD4 T cells from tnfaip3fl/fl Cd4-Cre or Tnfaip3fl/+ Cd4-Cre mice were stimulated with anti-CD3E and anti-CD28 antibodies, and Torin1 (25 nM) for 24 h. LC3 puncta were intracellularly stained with anti-LC3 antibody. Data are representative of 2 independent experiments. (B) The effect of Torin1 on ROS production in CD4 T cells. Naïve CD4 T cells from tnfaip3fl/fl Cd4-Cre or Tnfaip3fl/+ Cd4-Cre were stimulated as described in (A). CD4 T cells were stained with CM-H2DCFDA. Data are representative of 3 independent experiments. (C) Live cell proportions of Tnfaip3-deficient CD4 T cells in vitro. Naïve CD4 T cells were purified from congenically marked tnfaip3fl/fl Cd4-Cre and Tnfaip3+/+ Cd4-Cre mice and mixed at a 1:1 ratio. The mixed cells were stimulated with anti-CD3E and anti-CD28 antibodies, and Torin1 (25 nM) for 48 h in the same wells. Live cells were quantified by flow cytometry of DAPI-negative cells. Data are representative of 2 independent experiments. * p < 0.05 by the Student t test. Error bars indicate standard deviations.