Figure 7. GM-CSF triggers Zn binding to MTs in vivo and in human macrophages.

(A) Schematic, mice were left untreated or treated with rat IgG or anti-GM-CSF and infected i.n. with 2 × 10⁶ GFP⁺ yeasts (Hc); lungs were harvested 7 days p.i. and F4/80⁺CD11b⁺GFP⁺ and F4/80⁺CD11b⁺GFP⁻ cells were sorted. SEC-ICP-MS profiles of (B) F4/80⁺CD11b⁺GFP⁺ and F4/80⁺ CD11b⁺GFP⁻ cells isolated from lungs; (C) rat IgG treated and (D) anti-GM-CSF treated mice, 2 independent experiments; (E) SEC-ICP-MS profiles of lysates from human resting (h R) and GM-CSF activated (h Act) macrophages; Y axis, offset Zn signal; inset, zoom in of 15–25 min fraction, 2 independent experiments; (F) Dot plot of lung leukocytes gated on F4/80⁺CD11b⁺ macrophages; histogram of F4/80⁺CD11b⁺GFP⁺ macrophages; bar graphs are quantification of %, absolute numbers and MFI of F4/80⁺CD11b⁺GFP⁺ macrophages in rat IgG vs anti-GM-CSF treated mice, data are mean ± SEM, n=4; (G) Schematic representing transfer of CD45.1 WT macrophages into CD45.2 Csf2ra^−/− mice, infection and sorting of WT vs Csf2ra^−/− macrophages from lungs for gene expression analysis; (H) qRT-PCR of Mt1, Mt2 and Slc39a2 from sorted CD45.1⁺ (WT) and CD45.1⁻ (Csf2ra^−/−) GFP⁻ and GFP⁺ macrophages normalized to CD45.1⁺GFP⁻ macrophages, % values are decrease in gene expression in Csf2ra^−/− CD45.1⁻ GFP⁺ macrophages compared to WT CD45.1⁺ GFP⁺ macrophages, data are mean ± SEM, data are from 1 experiment. Related to Figure S6.