Figure S2.

Developmental analyses of monocytes and macrophages in FL and PND3 lungs and functional analyses of adult monocytes. (A–C) Flow cytometric analysis of immune cells derived from E19 FL isolated from Hem1^fl/flLysMCre⁺ and LysMCre⁺ control mice (A) and PND3 lungs isolated from WT and Hem1^−/− mice (B and C). (C) Total number of monocytes (Monos; singlet gated, ghost⁻CD45.2⁺CD11b^hiF4/80⁺CD11c⁻SiglecF⁻Ly6C⁺), fetal macrophages (MFs; singlet gated, ghost⁻CD45.2⁺CD11b^hiF4/80⁺CD11c⁻SiglecF⁻Ly6C⁻), pre-AMs (singlet gated, ghost⁻CD45.2⁺CD11b^intF4/80⁺CD11c⁺SiglecF⁻Ly6C⁻), and Mat-AMs (singlet gated, ghost⁻CD45.2⁺CD11b^intF4/80⁺CD11c⁺SiglecF⁺Ly6C⁻) harvested from PND3 lung tissues. (D) Pparγ (Pparg) expression as determined by real-time PCR in AMs harvested from 6–12-wk-old Hem1^fl/flLysMCre^+/+ and LysMCre^+/+ mice after culturing overnight in LPS (10 µg/ml). (E) Reduced BrdU incorporation by Hem-1–deficient AMs. Percentages of BrdU⁺CD45.2⁺CD11c⁺CD64⁺ cells in BALF from adult Hem1^fl/flLysMCre⁺ and LysMCre⁺ mice after 4 d of daily BrdU injections. Gating strategies are shown. Data shown are mean ± SEM from n = 4–9 mice/genotype representative of >2 independent exps. *, P < 0.05; **, P < 0.01; Student’s two-tailed t test. Rel., relative; SSC, side scatter.