Extended Data Fig. 3: IL-33 expands LT-expressing KLRG1+ ILC2s in PDAC mice.
From: IL-33-activated ILC2s induce tertiary lymphoid structures in pancreatic cancer
a, KLRG1+ ILC2 gating strategy. b, Lta and Ltb mRNA expression in purified KLRG1+ ILC2s (digital droplet PCR – left; scRNA-seq – right). c, d, f, LT expression on KLRG1+ ILC2s by flow cytometry (c, rIL-33-treated PDAC mice; f, rIL-33-treated DSS-colitis mice), and confocal imaging of purified intratumoural KLRG1+ ILC2s (d). In d, LT expression by flow cytometry (left) and confocal imaging (right) on αCD3 and αCD28-stimulated WT and Ltb−/− T cells (flow cytometry histograms) shown as positive and negative controls. e, Frequency of intratumoural KLRG1+ ILC2s in rIL-33-treated PDAC mice. PDAC 1–6 = orthotopic PDAC mice established with PDAC cell lines 1–6. g-i, scRNA-seq of 794 purified tumour ILC2s (g) and 1,668 purified tumour and draining lymph node (DLN) ILC2s (h, i) from PDAC mice. Heat map = top 25 genes for each cluster. UMAP, violin plots = ILC transcription factors (h), and markers (i) in unsupervised clusters in Fig. 3a, and ILC2 cytokines (h). Tbx21 was undetectable. j, ILC transcription factors on intratumoural KLRG1+ ILC2s from rIL-33-treated PDAC mice by flow cytometry. Data collected 10 days (g-j) and 2 weeks (b-f) after tumour implantation, pooled from ≥2 independent experiments with n ≥ 2 mice per group with consistent results. n = individual tumours from individual mice analysed separately. Horizontal bars = median; violin plots show the distribution with minima, maxima, and median (lines). P values by two-way ANOVA with Tukey’s multiple comparisons test (b left, e, f), Wilcoxon signed rank test (b, i violin plots), two-way ANOVA with Holm’s post-test (c), and two-way ANOVA with Sidak’s multiple comparisons test (d).
