Extended Data Fig. 8: Acute intranasal papain and IL-33 challenges in LCR1-deficient mice.
From: Multiscale 3D genome organization underlies ILC2 ontogenesis and allergic airway inflammation
a, Schematic representation of (left) the experimental acute papain or (right) recombinant IL-33 (rIL-33) challenges. Mice were challenged intranasally with either 30mg of papain (in 20mL of PBS) or with 200ng of rIL-33 (in 20ml of PBS) for four consecutive days. Euthanasia was carried out 24 hours after the last challenge. Lung parenchyma ILC2s and eosinophil numbers were assessed by flow cytometry. b, Flow cytometry gating strategy used for the identification of lung parenchyma ILC2s and eosinophils in LCR1+/+ and LCR1−/− mice challenged with papain or LCR1+/+ mice treated with PBS (20mL). Cells were pre-gated on SSC/FSC and Singlets. c, Quantification of ILC2 frequency among CD45+ cells, ILC2 and eosinophil numbers in LCR1+/+ and LCR1−/− mice challenged with papain, or LCR1+/+ mice treated with PBS (20mL). Data represents one experiment with males (green dots; n = 4 LCR1+/+ PBS, 4 LCR1+/+, and 4 LCR1−/− papain – 11.4-12.2-week-old). Error bars = SEM; and p-values: ns = not significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.0005, **** = p ≤ 0.0001 (One-way ANOVA with multiple comparisons and Bonferroni correction). d, Flow cytometry gating strategy used for the identification of lung parenchyma ILC2s and eosinophils in LCR1+/+ and LCR1−/− mice challenged with rIL-33 (200ng) or LCR1+/+ mice treated with PBS (20mL). Cells were pre-gated on SSC/FSC and Singlets. e, Quantification of ILC2 frequency among CD45+ cells, ILC2 and Eosinophil numbers in LCR1+/+ and LCR1−/− mice challenged with rIL-33 (200ng), or LCR1+/+ mice treated with PBS (20mL). Data is representative of two independent experiments (green dots; n = 3 LCR1+/+ PBS, 3 LCR1+/+ rIL-33, and 4 LCR1−/− rIL-33 – 9.4-9.9-week-old) and was repeated two times. Error bars = SEM; and p-values: ns = not significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.0005, **** = p ≤ 0.0001 (One-way ANOVA with multiple comparisons and Bonferroni correction).
