Figure 2.
The CD28-PI3K axis induces Zbtb32 expression after CD8+T cell activation. (A) The level of Zbtb32 expression was measured by RT-qPCR in activated CD8+ T cells in vitro (n = 3 for each group). (B) Quantifications of Zbtb32 expression levels were measured by RT-qPCR. The cells were activated by gradient dilutions of anti-CD3 plus fixed dilution of anti-CD28 or fixed dilution of anti-CD3 plus gradient dilutions of anti-CD28 (n = 3 for each group). (C) Quantifications of co-culture cell ratios of WT versus Zbtb32−/− CD8+ T cells were measured by flow cytometry. The cells were activated by gradient dilutions of anti-CD3 plus fixed dilution of anti-CD28 or fixed dilution of anti-CD3 plus gradient dilutions of anti-CD28 (n = 3 for each group). (D) The Zbtb32 expression level was measured by RT-qPCR in activated CD8+ T cells in the presence of various inhibitors or antibodies (n = 3 for each group). (E) The Zbtb32 expression level was measured by RT-qPCR in CD28-and Y189F mutant CD28 OE CD8+ T cells (n = 3 for each group). (F) Quantifications of live cell number and relative MFI of GzmB, IFNγ, and TNFa in CD28- and Y189F mutant CD28 OE WT and Zbtb32−/− activated CD8+ T cells were measured by flow cytometry. The levels in the Y189F mutant CD28 OE group were normalized (n = 5 for each group). Data in all graphs are shown as means ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by paired or unpaired two-tailed Student’s t test. Data shown are a representative of at least two independent experiments. Refer to the image caption for details.

The CD28-PI3K axis induces Zbtb32 expression after CD8 + T cell activation. (A) The level of Zbtb32 expression was measured by RT-qPCR in activated CD8+ T cells in vitro (n = 3 for each group). (B) Quantifications of Zbtb32 expression levels were measured by RT-qPCR. The cells were activated by gradient dilutions of anti-CD3 plus fixed dilution of anti-CD28 or fixed dilution of anti-CD3 plus gradient dilutions of anti-CD28 (n = 3 for each group). (C) Quantifications of co-culture cell ratios of WT versus Zbtb32−/− CD8+ T cells were measured by flow cytometry. The cells were activated by gradient dilutions of anti-CD3 plus fixed dilution of anti-CD28 or fixed dilution of anti-CD3 plus gradient dilutions of anti-CD28 (n = 3 for each group). (D) The Zbtb32 expression level was measured by RT-qPCR in activated CD8+ T cells in the presence of various inhibitors or antibodies (n = 3 for each group). (E) The Zbtb32 expression level was measured by RT-qPCR in CD28-and Y189F mutant CD28 OE CD8+ T cells (n = 3 for each group). (F) Quantifications of live cell number and relative MFI of GzmB, IFNγ, and TNFa in CD28- and Y189F mutant CD28 OE WT and Zbtb32−/− activated CD8+ T cells were measured by flow cytometry. The levels in the Y189F mutant CD28 OE group were normalized (n = 5 for each group). Data in all graphs are shown as means ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by paired or unpaired two-tailed Student’s t test. Data shown are a representative of at least two independent experiments.

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