Fig. 6: Two SFR members, namely SLAMF3 and SLAMF5, suppresses ILC2 activation in MLN.

a After 3 h of PMA plus ionomycin restimulation on day 6, the intracellular staining was performed to detect the expression of IL-5 and IL-13 in MLN ILC2s from mice following IL-33 treatment (i.n., day 0, 1 and 2). n = 4, 4, 5, 5, 4, 4, 5, 5, 4, 4, 6, 3 mice (left to right groups). b WT ILC2s were stimulated with OP9-DL1 cells ectopically expressing individual SFR member in the presence of IL-7 plus IL-33 for 72 h. Following a 3-h restimulation with PMA plus ionomycin, the expression of IL-5 and IL-13 by ILC2s was analyzed using intracellular staining. n = 5 per group. c Confocal microscopic analysis of the polarization of SLAMF3 or SLAMF5 (red) on the contact surface (yellow arrow) between ILC2s (KLRG1, sky blue) and their bystander T cells (CD3, green) within the MLN from mice on day 6 following IL-33 treatment (i.n., day 0, 1 and 2). d Representative flow cytometric profiles (left) and quantification (right) showing IL-5 and IL-13 expressing in gated MLN ILC2s after 3 h of PMA plus ionomycin restimulation on day 6 following IL-33 treatment (i.n., day 0, 1 and 2) in the indicated mice. n = 5, 6, 5 mice (left to right groups). The indicated mice were treated with IL-33 plus 2W1S peptide (i.n., day 0, 1 and 2), and the numbers of eosinophils (e), TH2 cells, CD69⁺ CD4⁺ T cells, and tetramer⁺ CD4⁺ T cells (f) were analyzed on day 9. n = 6, 6, 5, 6, 6, 4 (e) and n = 6, 5, 4, 6, 5, 4, 6, 5, 4 (f) mice in groups shown from left to right. The data represent two (a–c) or at least three (d–f) independent experiments with similar results. All data are represented as means ± SEM, and statistical analysis was conducted using a two-tailed Student’s t test (a) or one-way ANOVA (b, d–f). ns not significant. Please refer to Supplementary Fig. 6 for further details.