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. 2021 Aug 25;218(10):e20210745. doi: 10.1084/jem.20210745

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© 2021 Gschwend et al.

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Figure 2. — Hematopoietic-derived GM-CSF is dispensable for the development of AMs in the neonatal lung. (A–F) Analysis of P10 lungs isolated from Csf2^fl and Vav1^iCre/+;Csf2^fl mice. (A) Flow cytometry analysis of the Thy1.2⁺tdTomato⁺ population, gated on CD45⁺ cells. (B) Flow cytometry analysis of the EpCAM⁺ tdTomato⁺ population, gated on CD45⁻ cells. (C) Flow cytometry analysis of GM-CSF production in ILC2s (CD45⁺Lin⁻Thy1.2⁺ST2⁺) after restimulation with PMA/ionomycin (top row) or incubation with medium only (bottom row). (D) Percentage of GM-CSF⁺ ILC2s after restimulation or in the presence of medium only. (E) Flow cytometry analysis of CD11c⁺SiglecF⁺ AMs, gated on CD45⁺Ly-6G⁻ cells. (F) Quantification of AMs. (G) Quantification of AMs in P10 lungs of Rag2^+/+;Il2rg^+/+ and Rag2^−/−;Il2rg^−/− mice. (H) Quantification of AMs (CD45⁺SiglecF⁺CD11c⁺) in P10 lungs of Mcpt8^YFP-Cre;R26^+/+ and Mcpt8^YFP-Cre;R26^DTA/+ mice. Mcpt8^YFP-Cre/+ mice are indicated by circles, while Mcpt8^{YFP-Cre/YFP-Cre} mice are indicated by triangles. (I) Representative IF pictures of tdTomato⁺ cells (red) and CD45⁺ cells (green) in P10 lungs of Csf2^fl, Vav1^iCre/+;Csf2^fl, and Csf2^Δ/Δ mice (scale bars 20 µm). (A–C, E, and I) Data are from one experiment representative of at least two independent experiments. (D and F–H) Data are pooled from two (D), four (F), or three (G and H) independent experiments. ns, P ≥ 0.05; ****, P < 0.0001.