Extended Data Fig. 6: Excessive FAO suppresses ILC3 activation.
a, Representative Seahorse palmitate-FAO assay result showed the impact of palmitate (PA) on FAO of LTi (n = 5 per group). b, Basal OCR, ATP-linked OCR, maximal OCR and SRC were quantified (n = 3 per group; **p = 0.0097, **p = 0.0082, *p = 0.0163, *p = 0.0462). c, After a 3-day PA (20 μM) or vehicle treatment, IL-22 expression in activated LTi was assessed . Proportion of IL-22+ LTi and their IL-22 MFI were calculated (n = 4 per group; **p = 0.001, *p = 0.0117). d, Viability of LTi after PA (100 μM) treatment was assessed (n = 5 per group; ns). e, CPT1A overexpression in LTi was measured by qRT-PCR (n = 5 per group; *p = 0.0297). f, Impact of CPT1A on IL-22 production in LTi were assessed. Proportion of IL-22+ LTi and their IL-22 MFI were calculated (n = 5 per group; *p = 0.0436, ***p < 0.0001). g, After a 3-day PA or vehicle treatment, IL-22 production in activated NKp46+ ILC3 was assessed. Proportion of IL-22+ NKp46+ ILC3 and their IL-22 MFI were calculated (n = 5 per group; ***p < 0.0001, ***p < 0.0001). (h and i) Rag2-/- mice were fed with HFD (high fat diet) or CD (chow diet) for 5 weeks. IL-22 expression in siLP NKp46+ ILC3 (h) and colonic LTi (i) was examined. Proportion of IL-22+ siLP NKp46+ ILC3 and their IL-22 MFI (n = 4 per group; *p = 0.0269, *p = 0.0151)(h), and proportion, IL-22 MFI and number of IL-22+ colonic LTi (n = 4 per group; *p = 0.0298, *p = 0.0477, *p = 0.0496)(i), were calculated. (j and k) Enriched features in PA untreated (j) and treated (k) LTi was analyzed using two-sided GSEA adopted permutation-test. Nominal p-value and NES were calculated. Data are representative of at least three independent experiments (a-i) or two independent experiments (j-k). Data are presented as the mean ± s.d., and statistical significance was determined by two-sided unpaired t-test (b-i). Ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001.
