DCAF13-deficient T cells showed normal TCR response and activation. (A) Representative flow cytometry pictures (upper) and corresponding quantification (bottom) of the DN, DP, CD4SP, and CD8SP thymocyte subpopulations from OT1-TG CD4 Cre⁻Dcaf13^fl/fl (WT) and OT1-TG CD4 Cre⁺Dcaf13^fl/fl (cKO; WT, n = 6; cKO, n = 3). (B) Representative flow cytometry pictures (left) of the percentage of CD4⁺ and CD8⁺ T cell subsets in lymph nodes and spleen from OT1-TG CD4 Cre⁻Dcaf13^fl/fl (WT) and OT1-TG CD4 Cre⁺Dcaf13^fl/fl (cKO) and corresponding quantitation (right; WT, n = 6; cKO, n = 3). (C) The phosphorylation level of PLC-γ, AKT, and ERK in T cells from CD4 Cre⁻Dcaf13^fl/fl (WT) and CD4 Cre⁺Dcaf13^fl/fl (cKO) mice after stimulation with anti-CD3/CD28. Quantification of phosphorylation level in total targeted proteins with ImageJ. (D and E) Flow cytometry analysis of CD69 (D) and CD25 (E) in naïve CD4⁺ T cells isolated from CD4 Cre⁻Dcaf13^fl/fl (WT) and CD4 Cre⁺Dcaf13^fl/fl (cKO) mice under indicated anti-CD3/CD28 stimulation for 4 h. Representative flow cytometry pictures (left) and corresponding quantitation (right; n = 3) are shown. (F and G) Flow cytometry analysis of CD69 (F) and CD25 (G) in naïve CD8⁺ T cells isolated from CD4 Cre⁻Dcaf13^fl/fl (WT) and CD4 Cre⁺Dcaf13^fl/fl (cKO) mice under indicated anti-CD3/CD28 stimulation for 4 h. Representative flow cytometry pictures (left) and corresponding quantitation (right) (n = 3) are shown. Data are presented as the mean ± SD. A two-tailed Student’s t test was used. ns is not significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are available for this figure: SourceData F3.