Fig 2: mRNA and protein expression of key functional molecules of ILC2s. (A–C) ILC2s were marked as CD45+Lin-CD127+CRTH2+ and sorted via FACS. (A) A heatmap showing representative results of mRNA expression of the key functional molecules of ILC2s obtained from HD-PB, LU-PB, and LU-CA samples. (B, C) The relative mRNA expressions of IL4 (B) and IL13 (C) in ILC2s from HD-PB, LU-PB, and LU-CA samples (for each group, n=18). (D–G) Protein expression of IL-4 (D, E) and IL-13 (F, G) in ILC2s from HD-PB, LU-PB, and LU-CA samples assessed via flow cytometry (for each group, n=20). HD-PB: PBMCs from healthy donors, LU-PB: PBMCs from NSCLC patients, and LU-CA: tumor single cell suspension from NSCLC patients. NS, not significant; *P<0.05, ***P<0.001, ****P<0.0001. P values were calculated by one-way ANOVA and post hoc comparisons were performed via Tukey’s test.

Fig 3: PD-1high ILC2s expressed higher levels of IL-4 and IL-13. PD-1high ILC2s and PD-1low ILC2s were marked as CD45+Lin-CD127+CRTH2+PD-1high and CD45+Lin-CD127+CRTH2+ PD-1low, respectively, and sorted by FACS. (A, B) The gene expressions of PDCD1 (encoding PD-1) (A), IL4, and IL13 (B) were upregulated in the PD-1high group compared with the matched PD-1low group (n=26). (C–F) The representative results and statistical results of IL-4 (C, D) and IL-13 (E, F) protein expression according to the different PD-1 levels assessed by flow cytometry (for each group, n=26). Each dot represents one sample. ***P<0.001, ****P<0.0001. P values were calculated using a paired two-tailed student’s t-test.

Fig 4: Genome-wide transcriptomic analysis of av-deficient TH cells(a) Principal component analysis of the transcriptomes of control versus av-deficient TH cells. (b) Volcano plot depiction of differentially expressed genes in control versus av-deficient TH2 cells. (c) & (d) RNA expression of type 2 genes by control or av-deficient TH cells; Il13 ***P=0.0002, **P=0.0012; Il5 ***P=0.0006, **P=0.0025; Il4 **P=0.0046, *P=0.0167, Gata3 ****P<0.0001. (e) RNA expression of type-1 signalling genes by control or av-deficient TH cells; Statl ****P<0.0001, *P=0.0438; Isg2O *P=0.0148; Runx3 **P=0.0052, *P=0.0220. (c) - (e) mean ± SEM; one-way ANOVA with Tukey’s post-hoc test. (f) Flow cytometric analysis of Runx3 expression by av-deficient TH cells. Data are representative of 2 independent experiments; unpaired two-sided t-test; ***P=0.0003. (g) KEGG pathway analysis of genes downregulated in av-deficient versus control TH2 cells.