CyTOF analysis of thymocytes from WT and STING N153S mice. Thymocytes were harvested from 4–6-mo-old WT and STING N153S mice and analyzed by CyTOF. (A) viSNE map of representative WT and STING N153S thymocyte populations illustrating color-coded cell populations that were clustered together based on similarity in cell surface marker expression. (B) Contour plots of viSNE maps illustrating the density of cell populations as defined in A. (C–M) The total number of thymocytes was determined. The percentage of thymocytes labeled with each of the respective surface markers was measured by CyTOF and used to calculate the total numbers of CD45⁺ cells (C). Percentages of other populations are displayed as follows: CD4⁻CD8⁻ (double negative; D), CD4⁺CD8⁺ (double positive; E), CD4⁺ (F), CD8⁺ (G), CD19⁺ (H), NK1.1⁺(I), CD25⁻CD44⁺ (DN1; J), CD25⁺CD44⁺ (DN2; K), CD25⁺CD44⁻ (DN3; L), and CD25⁻CD44⁻ (DN4; M) cells. (N) Percentage Thy1.1⁻ WT littermate or STING N153S double-negative thymocytes from mixed bone marrow chimeras. Data in C–M represent the mean of samples from n = 6 mice per genotype from two independent experiments analyzed by Mann-Whitney test. **, P < 0.005. Data in N represent the mean of samples from n = 3 bone marrow chimeric mice analyzed by unpaired t test. ****, P < 0.0001.