Fig. 4: Differential CD69 and CD103 expressions defined silicotic CD4⁺ T_RM cells into effector or regulatory T_RM cells.

a Flow plot indicated CD4⁺ T_RM cells in fibrotic lungs were divided into four subsets by CD69 and CD103 expressions. CD69⁺CD103⁺ (Red); CD69⁺CD103^– (Blue); CD69^–CD103⁺ (Orange); CD69^–CD103^– (Green). b–d Flow histogram displayed transcriptional factor T-bet (b), ROR-γt (c), and GATA-3 (d) expressions among different subsets. Bar graph below shows the MFI among the indicated CD4⁺ T_RM cell subsets. e Flow histogram compared ratios of FOXP3⁺ among different subpopulations. Bar graph below reveals the percentages of FOXP3⁺ among the indicated subsets. f FC heatmaps show the expression intensity of each marker. Representative flow histogram compared the intensity of CD25, PD-1, ST2, ICOS, and CD39 among four subsets in CD4⁺ T_RM cells. g Typical flow plots showed CD69 and CD103 expressing patterns on Tregs (FOXP3⁺) or Teff cells (FOXP3^–) in CD4⁺ T_RM cells. h Typical flow plots demonstrated CD69 and CD103 expressing patterns on T-bet⁺, ROR-γt⁺, or GATA-3⁺ subsets in CD4⁺ T_RM cells. i The bar graph showed the ratios of CD103⁺ against CD103^– subsets in CD69⁺ CD4⁺ T_RM cells. Bar graphs are the combined results of at least three independent experiments. Individual mice are plotted on the graphs, n = 5–6 biologically independent animals. Values are reported as the mean ± SD. P value is determined by one-way ANOVA followed by Tukey’s test.