Panel A shows flow cytometry plots of CD4-positive Foxp3-positive, CD4-positive Foxp3-negative, and CD8-positive T-cell populations from spleen samples. Panel B shows histology images of skin, lung, liver, kidney, stomach, small intestine, and colon tissues from wild-type and knockout mice. Panels C and D show flow cytometry plots of CD8-positive and CD4-positive T-cell subsets, including CD127 and Foxp3 expression profiles, from spleen samples. Panel E shows bar plots with x-axis Experimental group and y-axis Percentage of CD8-positive T cells or CD8-positive T-cell subset percentages. Panel F shows bar plots with x-axis Experimental group and y-axis Percentage of CD4-positive T cells or CD4-positive T-cell subset percentages. Panel G shows bar plots with x-axis Experimental group and y-axis Percentage of CD8-positive T cells or CD8-positive T-cell subset percentages in the lacrimal gland. Panel H shows bar plots with x-axis Experimental group and y-axis Percentage of CD4-positive Foxp3-positive T cells or subset percentages in the lacrimal gland. Panel I shows bar plots with x-axis Experimental group and y-axis Percentage of CD8-positive T cells or CD8-positive T-cell subset percentages in the salivary gland. Panel J shows bar plots with x-axis Experimental group and y-axis Percentage of CD4-positive Foxp3-positive T cells or subset percentages in the salivary gland.
Characterization of Cre + -Tnfrsf9 +/+ and Cre + -Tnfrsf9 fl/fl mice, related to Fig. 1. (A) CD137 expression on splenic Foxp3+ and Foxp3− CD4 T cells and CD8 T cells of Cre+-Tnfrsf9+/+ and Cre+-Tnfrsf9fl/fl mice. NOD.Tnfrsf9−/− cells were used as the negative control. Representative flow cytometry profiles of three mice per genotype are shown. (B) Representative histological images of indicated tissues of three prediabetic 10- to 11-wk-old female mice per genotype are shown. The scale bar is 1 mm. Similar results were observed in three age-matched male mice per genotype. (C and D) Representative flow cytometry plots of splenic CD8 (C) and CD4 (D) T cells in 6- to 8-wk-old Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) females. The CD3 by CD4 and CD25 by Foxp3 plots of the KO spleen are also shown in Fig. 1 A. (E) Percentages of splenic and PLN total CD8 T cells in 6- to 8-wk-old Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) females and their activation status determined by the expression levels of CD44 and CD127. Summarized results from three experiments are shown. *P < 0.05 by an unpaired t test. ns: not significant. (F) Percentages of splenic and PLN total CD4 T cells and the activation status of Foxp3− CD4 T cells in 6- to 8-wk-old Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) females determined by the levels of CD44 and CD127 expression. Summarized results from three experiments are shown. ns: not significant by an unpaired t test. (G and H) Percentages of total CD8 (G) and CD4 (H) T cells and their activation status in lacrimal glands of 9- to 11-wk-old Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) male mice. Results are summarized from three experiments. **P < 0.005 by an unpaired t test. ns: not significant. (I and J) Percentages of total CD8 (I) and CD4 (J) T cells and their activation status in salivary glands of 9- to 10-wk-old Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) female mice. Results are summarized from two experiments. *P < 0.05 by an unpaired t test. ns: not significant.