Fig 2: CD206+ ATMs are distinct from other subtypes.(A) Dendrogram based on the top 500 variably expressed genes (left) and PCA (right) of human ATM subtypes. The 3 ATM subtypes were each isolated from 3 patient samples with obesity (2 DM, 1 NDM) for a total of 9 analyzed cell populations. (B) Distance diagram between ATM subtypes based on DE genes for each comparison. (C) Top pathways distinguishing ATM subtypes. (D) Scavenger receptor–related gene expression in ATM subtypes. (E) Overlap of ATM subtypes with Vijay et al. data (13). Fold difference in expression for each comparison is shown, with gray bars showing expression in CD206+ ATMs, dark blue in CD11c+ ATMs, and light blue in DP ATMs. Overlap was defined as a higher percentage of mutual genes than the overall mean percentage of mutual genes across all 5 Vijay populations. (F) Overlap of ATM subtypes with Wentworth et al. data (9). (G) Overlap of ATM subtypes with monocytes in obesity (GSE32575). (H) MacSpectrum analysis of ATM subtypes. MPI, macrophage polarization index; AMDI, activation induced differentiation index.

Fig 3: Depletion of APP altered surface expression of M0 (a) and M2c (f). Combined depletion of APP and ELAVL-1 in iPSC-CM increased CD206+ cells in M1 (b), M2 (c) and M2a (d) subtype. Depletion of APP in the iPSC-CM reduced CD163+/CD206+/CD80+ populations in M0 subtype compared to control. APP depletion increased CD163-/CD206+/CD80- populations in M2c subtype iPSC-CM treated macrophages (p < 0.05) and compared to control media (p < 0.05). (n = 4; Mean ± SEM). Finally, depleting both APP and ELAVL-1 in iPSC-CM led to increased expression of CD206 in M1, M2, and M2a subtypes. In M2c subtype, the CD206+ population was also increased, but only compared to control cells. In M2a subtype (e), an additional subpopulation of (CD163-/CD206+/CD80+) was increased. (n = 4; Mean ± SEM). Values are represented in Supplementary Materials Table S1. Flow cytometry histograms are represented as Supplementary Figures S1 and S2.

Fig 4: ATM subtype cytokine secretion and localization.(A) Expression of cytokines in NDM and DM obese whole visceral adipose tissue. Data are shown as mean FPKM log2-fold difference. (B) Expression of cytokines in CD206+ and CD11c+ ATMs by RNA-Seq. (C) Baseline cytokine secretion from sorted CD206+ or CD206- VAT ATMs, shown as mean concentration. (D) Cytokine secretion from sorted, LPS-stimulated CD206+ or CD206- VAT ATMs. Each replicate was normalized to baseline, and measures are shown as mean fold over baseline ± SD. (C and D) Replicates: n = 3–4 wells per measure. Raw data were analyzed by 2-way ANOVA with Sidak’s multiple comparison test. (E) Immunostaining for CD206 (green) and CD11c (red) in 2 human visceral adipose tissue samples in obesity. CD206 and CD11c show differential expression in CLS (white arrows) and FALC (orange arrow), but they show coexpress in cells between adipocytes (blue arrows). Blood vessels (blue) were stained with UEA-1 lectin. Scale bar: 100 µm. Row 1: epifluorescence microscopy. Rows 2–4: confocal microscopy.

Fig 5: Glufosinate treatment promotes M1-like TAMs, CTL tumor accumulation, vessel pruning, and normalization and inhibits metastasis in a murine model of melanoma AExperimental plan. Mice were intradermally injected with YUMM 1.7 (CD90.1+) cells. When the tumor reached 80–100 mm3 size, mice were treated by gavage with glufosinate (20 mg/kg) or vehicle for 14 days.B, CIntradermal YUMM 1.7 (CD90.1+) tumor growth over the time (B) and number of metastatic lesions per lung (C) in vehicle and glufosinate (20 mg/kg)-treated mice (pool of 2 independent experiments; 10 mice per condition in total).DRepresentative images of metastatic lesions checked by DAB-H stain for CD90.1 positivity in vehicle and glufosinate (20 mg/kg)-treated mice (n = 6). Six images per lung were analyzed. Scale bar: 2 mm. Zoomed area scale bar: 50 µm.E, FFACS quantification of M1-like MHC class IIhigh (E) and MHC class IIlow (F) CD11c+ CD206- TAMs over the total number of F4/80+ cells in vehicle and glufosinate (20 mg/kg)-treated mice (n = 6).GQuantification and representative images of CD11c+ cells over the total number of F4/80+ cells in vehicle and glufosinate (20 mg/kg)-treated mice (n = 6). Six images per tumor were analyzed. Scale bars: 50 µm.H, IFACS quantification of M2-like MHC class IIhigh (H) and MHC class IIlow (I) CD11c- CD206+ TAMs over the total number of F4/80+ cells in vehicle and glufosinate (20 mg/kg)-treated mice (n = 6).JQuantification and representative images of CD206+ cells over the total number of F4/80+ cells in vehicle and glufosinate (20 mg/kg)-treated mice (n = 6). Six images per tumor were analyzed. Scale bars: 50 µm.K, LFACS quantification of M1-like MHC class IIhigh (K) and M2-like MHC class IIlow (L) TAMs over the total number of F4/80+ cells in vehicle and glufosinate (20 mg/kg)-treated mice (n = 6).MQuantification of MHC-II protein, expressed as mean fluorescence intensity (MFI), on F4/80+ cells, was obtained by flow cytometry (n = 6).NRT–PCR quantification of M2 (Arg1 and Cxcr4) and M1 (Cxcl9 and Nos2) markers in vehicle and glufosinate (20 mg/kg)-treated mice (n = 6).Data are reported as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 ****P < 0.0001.Exact P values and statistical tests are reported for each experiment in Appendix Table S2. Source data are available online for this figure.