Fig. 7: SLAMF3 and SLAMF5 inhibit the downstream NF-κB pathway of IL-7/IL-33 through SHIP-1.

a Detection of SAP expression in the indicated cells on day 9 following IL-33 treatment (i.n., day 0–2). b Detection of IL-5 and IL-13 expression in MLN ILC2s on day 6 following IL-33 treatment (i.n., day 0–2). n(WT) = 3 and n(SAP^−/−EAT-2^−/−) = 4 mice. c Detection of SHP-1, SHP-2, and SHIP-1 expression in MLN ILC2s on day 6 following IL-33 treatment (i.n., day 0–2). Isotype, fluorescently labeled secondary antibody, representing negative control. Detecting of SHP-1 (d), SHP-2 (e), and SHIP-1 (f) phosphorylation in MLN WT (Blue line) and SLAMF3/5^−/− (red line) ILC2s on day 6 following IL-33 treatment (i.n., day 0–2). Gray shade, isotype. n = 5, 9, 7 (d) or 3, 3, 3 (e) or 8, 12, 10 (f) mice (left to right groups). g Western blot analysis of the phosphorylated SHIP-1 (P-SHIP-1) in WT ILC2s with antibody-mediated crosslinking of SLAMF3 (top) and SLAMF5 (bottom) at the indicated minute points (min). h Detection of IL-5 and IL-13 expression in MLN ILC2s of the CHIME chimera (refer to the methods) on day 6 following IL-33 treatment (i.n., day 0 to 2). CD90.1-negative: representing sgRNA-unexpressed control; CD90.1-positive: representing SHIP-1 sgRNA-expressed. n = 4 mice per group. i Representative overlaid histograms and quantification of Iκβ-α expression in MLN WT (Blue line) and SLAMF3/5^−/− (red line) ILC2s on day 6 following IL-33 treatment (i.n., day 0 to 2). Gray shade, representing isotype. n(WT) = 9 and n(SLAMF3/5^−/−) = 11 mice. Representative images (j) and quantification (k) of the phosphorylation of NF-κB in MLN ILC2s from the indicated mice on day 6 following IL-33 treatment (i.n., day 0 to 2). n(WT) = 40 and n(SLAMF3/5^−/−) = 70 cells. l, m Western blot analysis of NF-κB activation in WT ILC2s stimulated with (+) or without (−) antibodies targeting IL-7Rα (l) or IL-33R (m), in the presence (+) or absence (−) of antibodies against SLAMF3 or SLAMF5. The data represent at least two independent experiments with similar results. All data are represented as means ± SEM, and statistical analysis was conducted using two-tailed Student’s t test (b, h, i, k) or one-way ANOVA (d–f). ns not significant. Please refer to Supplementary Fig. 7 for further details.