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. 2021 Jun 2;218(7):e20201354. doi: 10.1084/jem.20201354

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© 2021 Verma et al.

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Figure 5. — scRNA-seq of lung hematopoietic cells. (A) A schematic showing study design and timeline for scRNA of lung cells from the memory model. (B) A t-SNE feature plot showing clustering of all CD45⁺ lung cells with mean expression of the WGCNA blue module, which contains genes characteristic ILC2s. The ILC2 cluster (circled) expressed Il7r, Gata3, Il1rl1, and Icos (violin plots) and Rora and Nfkb1 (feature plots). (C) Log fold-change (LFC) of selected top 16 genes from the memory group 1B versus 1A (after and before recall) and Group1 (memory) versus Group 2 (Sal control). (D) qPCR analysis of mRNA expression of select genes identified by scRNA-seq in sorted ILCs from the lung tissue obtained from the study groups. *, P < 0.01; **, P < 0.001; ***, P < 0.0001, one-way ANOVA, n = 5/group. (E) FCM analysis of expression of CD44 and Thy1 on ICOS⁺ST2⁺ ILCs (CD45⁺CD25⁺ cells); and the expression of FHL2⁺, NR4A2 MPP7, and CCR8 in ICOS⁺ ILCs (CD45⁺CD25⁺) in the lung before and after recall. *, P < 0.01; **, P < 0.001; ***, P < 0.0001, n = 4–5/group, one-way ANOVA. Gr., group; Sac, sacrifice.