(A) Thymic CD4⁺ Tregs from KO and WT mice (n = 3) were screened for the mRNA expression of members in the ADAM family, and cumulative data (mean ± SEM) from 3 experiments are shown. (B) The mRNA expression of ADAM10 relative to β-actin in thymic CD4⁺ Tregs from WT mice (n = 3) that were incubated with the PP2A activator FTY720 (5 μM) or with the PP2A inhibitor OA (4 nM) for 4 hours. (C) The levels of soluble IL-2Rβ in the supernatants of thymic CD4⁺ Tregs from KO mice (n = 3) in the presence of the selective inhibitor ADAM10 (GI254023X) in different doses. (D) Representative flow cytometry histograms depicting the surface IL-2Rβ expression in thymic CD4⁺ Tregs from KO mice (n = 3) at baseline (accompanied by a dotted-line histogram representing the negative control) or in response to ADAM10 inhibitor (GI254023X; 10 μM; broken-line histogram), IL-2 (20 nM; dark-gray histogram), or their combination (light-gray histogram). Cumulative data (mean ± SEM) from 3 experiments are shown. (E) Soluble IL-2Rβ in the supernatants of thymic CD4⁺ Tregs from WT mice (n = 3) that were incubated with PP2A inhibitor OA (4 nM), ADAM10 inhibitor (GI254023X; 10 μM), or their combination. Cumulative data (mean ± SEM) from 3 experiments are shown. (F) Expression of p-STAT5 following ADAM10 inhibition (GI254023X; 10 μM) and IL-2 stimulation (20 nM) in thymic Tregs from WT (n = 3; dark-gray histogram) and KO (n = 3; light-gray histogram) mice. Cumulative data (mean ± SEM) from 3 experiments are shown.