Figure 4. M-CSF is essential to regulate mTORC2 signaling in M2 macrophages.

(A) Expression of PD-L2 and RELMα in macrophages cultured for 24 hr with (M2) or without (M0) IL-4 or M-CSF. (B) Expression of iNOS and TNF-α in macrophages cultured for 24 hr with (M1) or without (M0) IFN-γ + LPS or M-CSF. (C) Glucose consumption, and (D) basal ECAR and basal OCR of M2 macrophages treated as in (A). (E) Level of phosphorylated AKT^s473 in macrophages as in A, assessed by flow cytometry. MFI values are shown. (F) Scheme to examine the effect of blocking M-CSF/CSF1R interaction on M2 activation in vivo. (G) Phosphorylated NDRG1 (p-NDRG1) in pMacs was measured by immunoblot; band density was normalized to loading controls and is presented in arbitrary units (AU). (H) Level of AKT^s473, phosphorylation, and (I) frequency of RELMα⁺ cells, in pMacs. Data in A,B, E, H and I are from, flow cytometry and in A, B and E are from one experiment representative, but numbers represent mean % (A) or MFI (B,E) values, ± s.e.m, from three independent experiments. In C,D and G–I, data are mean ± s.e.m. of technical replicates from one experiment representative of three or more independent experiments. *P < 0.05, **P < 0.005 and ***P < 0.0001.