Supplementary Figure 8: SAP-Fyn and SHP-1 differentially influence iNKT cell development.
From: SLAM receptors foster iNKT cell development by reducing TCR signal strength after positive selection
(a) A schematic of SLAMF6 isoform 2 (SLAMF6–2) showing the four tyrosines in the cytoplasmic domain and the tyrosine (Y)-to-phenylalanine (F) mutants. Signal peptide (SP); extracellular domain (ECD); transmembrane domain (TM); cytoplasmic domain (CT); immunoreceptor tyrosine-based switch motifs (ITSM); Y295,319F (Y1,2F) and Y335,349F (Y3,4F). (b) Representative experiments of the data depicted in Fig. 8b. (c-e) Flow cytometry analyses showing various stages of iNKT cell development (c), PD-1 (d) and Bcl-2 (e) expression in the thymus and spleen of WT and SAPR78A KI mice. n = 4 (c); n = 6 (d); n = 7 (e). (f) Cytokine production in response to α-GalCer was evaluated as detailed in the legend of Fig. 3a. n = 3. (g) Flow cytometry analyses showing frequencies of thymic and splenic iNKT cells from hCD2-icre, WT, and SHP-1 cKO mice. n = 7. (h) Quantitative RT-PCR analysis showing Nur77-encoding RNA (Nr4a1) expression in iNKT cells from WT, SFR KO, WT (Ptpn6fl/fl) and SHP-1 cKO mice. n = 3. Results for SFR KO, WT (Ptpn6fl/fl) and SHP-1 cKO are normalized to WT. In (c-g), a representative experiment is shown on the left, while data from multiple mice are depicted on the right. Each symbol represents an individual mouse; error bars represent mean with s.d.. * P ≤ 0.05, *** P ≤ 0.001, ns P > 0.05 (two-sided unpaired t-test; two-sided paired t-test for f). Data are representative of the numbers of experiments as described for Fig. 8b (b); 3 (c); 4 (d); 2 (e, f); 3 (hCD2-icre) and 5 (WT and SHP-1 cKO) (g); and 3 (h) independent experiments.
