Fig 1: Characterization and gene profiling of AlloHSC-iNKT cells(A) FACS detection of surface markers on AlloHSC-iNKT cells. PBMC-iNKT and PBMC-Tc cells were included as controls.(B and C) Antigen responses of AlloHSC-iNKT cells. AlloHSC-iNKT cells were cultured for 7 days, in the presence or absence of aGC (denoted as aGC or Vehicle, respectively). (B) Cell growth curve (n = 3). (C) ELISA analyses of cytokine (IFN-?, TNF-a, IL-2, IL-4 and IL-17) production at day 7 post aGC stimulation (n = 3).(D) FACS detection of intracellular cytokines and cytotoxic molecules in AlloHSC-iNKT cells. PBMC-iNKT and PBMC-Tc cells were included as controls.(E–I) Deep RNA-seq analysis of AlloHSC-iNKT cells generated from CB- or PBSC-derived CD34+ HSCs (n = 3 for each). Healthy donor PBMC-derived conventional CD4- aß T (PBMC-aßTc; n = 8), CD4- iNKT (PBMC-iNKT; n = 3), ?d T (PBMC- ?dT; n = 6), and NK (PBMC-NK; n = 2) cells were included as controls. N indicates different donors. (E) Principal-component analysis (PCA) plot showing the ordination of all six cell types. (F–I) Heatmaps showing the expression of selected genes encoding transcription factors (F), NK activating and inhibitory receptors (G), tissue inflammatory homing markers (H), and HLA molecules (I) for all six cell types.Representative of 1 (E–I) and 3 (A–D) experiments. Data are presented as the mean ± SEM ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, by Student’s t test.