Figure 4.

Th2-like Tregs Exhibit Higher Chemotaxis toward CCL17/22 Than Other Th-like Tregs and Th2-like Teffs

(A and B) Volcano plots showing RNA-seq data obtained from activated Th-like Treg subsets (A), and heatmaps showing upregulation of leukocyte transendothelial migration, chemokines, and chemokine receptors genes between Th-like Treg subsets using Partek software and the KEGG database (B).

(C) Representative dot plots and percentage of migrated memory Teffs and Tregs. Memory Teffs (5 × 10⁴) and memory Tregs (5 × 10⁴) were placed in the top chamber of a 5-μm-pore Transwell filter system with ICAM (1 μg/mL). Bottom chambers were filled with media only; CCL17/22 (0.5 μg/mL), CCL20 (0.5 μg/mL), or CXCL10 (0.5 μg/mL); or a combination of all of them. The percentage of migration for each subset was calculated as (number of cells in the bottom chamber after 1 hr × 100)/initial number of cells in the top chamber.

(D and E) Representative dot plots and percentage of migration between Th-like Treg subsets (D) and between CCR4⁺ Th-like Teff and Th-like Tregs (E) (n = 6, mean ± SEM using bar charts, RM two-way ANOVA with Sidak’s test).

For all statistical tests, ^∗∗∗∗p < 0.0001, ^∗∗∗p < 0.001, ^∗∗p < 0.01, and ^∗p < 0.05 were considered significant.