Fig. 1: Hypoxia increases ILC3 activation and proliferation.

A Gating strategy of live Lin^-CD45^lowCD90.2^high enriched ILC3 population from small intestine (si) lamina propria (as shown in Fig. S1). B RORγt mean fluorescence intensity and production of IL-17 and IL-22 by enriched si-ILC3 subset obtained after incubation for 3 h, under normoxia/hypoxia. Cells were non-stimulated (mock) or stimulated with IL-1β + IL-23 ex vivo (n = 6). C Relative Rorc, Il22 and Il17 mRNA expression in stimulated MNK3 cells under normoxia/hypoxia (n = 4). D Cellular viability of MNK3 cells incubated 3 h in hypoxia/normoxia using annexin-V/7-AAD staining. Live (green) represents the double-negative population, early apoptosis (yellow) shows annexin-V⁺7-AAD⁻, and late apoptosis (red) is double positive cells (n = 4). Percentage of EdU⁺ MNK3 cells (E) and Ki67⁺ cells in the ILC3-enriched population (F) as measured by flow cytometry (n = 7 and 3, respectively). Representative EdU⁺ plots are presented on the left. Results are representative of at least two independent experiments and presented as mean ± SEM. *p < 0.05; **p < 0.01.