Fig 1: Liver sinusoidal endothelial cells supported the differentiation of liver-type ILC1s(A) Sorted peripheral blood ILCPs (Figure S5A; Lin-CD45+CD127+CD94-NKp80-CD45RA+CD117+CD294-KLRG1-NKP44-) from four healthy donors were cultured for 2 weeks on OP9-DL4 feeder cells or LSECs with IL-7 +/- TGF-ß1 as shown to the left. The dot plots to the right show the expression patterns of CD56 and CD94 among Lin- lymphocytes derived in the indicated culture conditions.(B) Frequencies of CD200R1+CD49a+EOMES- cells among total Lin-CD45+CD94+CD56+ cells derived in vitro following culture of ILCPs (n = 8) as in (A).(C) Representative flow cytometry analyses of EOMES, T-BET, perforin, CXCR6, NKp80, CD200R1, and CD49a expression by Lin-CD45+CD94+CD56+ cells derived from ILCPs in the indicated culture conditions. Fluorescence minus one (FMO) control with the respective lacking specific antibody is shown as black dotted lines in each histogram.(D) Intracellular flow cytometry analysis of IFN-?, IL-2, GM-CSF, TNF-a, and IL-22 production by in vitro derived Lin-CD45+CD94+CD56+ cells generated from ILCPs as in (A) and following stimulation for 5 h with PMA and ionomycin. Unstimulated cells were set as controls (dotted black line).(E) Representative flow cytometry analyses showing expression of the indicated markers by total Lin-CD45+ cells obtained after 2 weeks culture of primary sorted liver-type hepatic cNK, CD49a+trNK, or CD49a-trNK cells on OP9-DL4 feeder cells or LSECs with IL-7 +/- TGF-ß1 as shown to the left. Data are representative of at least three independent experiments.(F) Representative flow cytometry analyses showing expression of the indicated markers by total Lin-CD45+ cells obtained after 2-week culture of primary sorted liver-type ILC1s (LT-ILC1s) or hepatic NK cells (sorted as Lin-CD56+CD94+NKp80+) with OP9-DL4 cells and IL-7. Data are representative of at least three independent experiments.(G) Frequencies of CD200R1+CD49a+EOMES-cells among total Lin-CD45+CD94+ lymphocytes obtained from cultures initiated with either total hepatic NK cells or liver-type ILC1s (F).*p < 0.05, **p < 0.01, ***p < 0.001, error bars represent SEM. See Figure S6.