FIG. 1.

CD11c⁺ eATMΦs exhibiting differential MGL1 expression (MGL1^med/CD11c⁺ and MGL1⁻/CD11c⁺) accumulate in the eAT of mice fed an HFD. SVCs were obtained by collagenase digestion from eAT of mice fed an HFD or LFD for 8 weeks, labeled with F4/80, MGL1, and CD11c antibodies and analyzed by flow cytometry. A: HFD-associated increase in F4/80⁺/CD11c⁺ eATMΦs reflects increases in two ATMΦ subtypes, designated MGL1^med/CD11c⁺ (R2) and MGL1⁻/CD11c⁺ (R1) according to their MGL1 staining intensity. B: Confirmation of MGL1 protein expression in MGL1⁺/CD11c⁻ and MGL1^med/CD11c⁺ eATMΦs by fluorescence microscopy of sorted eATMΦs (as in A). C: Gene expression for F4/80, MGL1, and CD11c in sorted eATMΦ subtypes. D: Quantification of eATMΦ subtypes in response to 8 and 12 weeks of HFD demonstrating the absolute and proportional increase in CD11c⁻ and CD11c⁺ subtypes during the HFD time course. *P < 0.05; **P < 0.01, ANOVA and Tukey test. E: Evidence that MGL1^med/CD11c⁺ eATMΦs are recruited rather than derived by phenotypic progression from resident MGL1⁺/CD11c⁻ cells. Lean mice were pulsed with the phagocyte-specific dye PKH26 and subsequently fed 2 standard diet or an HFD for 1 month followed by FACS of eATMΦs. Only ATMΦs present in eAT before clearance of the dye (24 h) express the label. (A high-quality digital representation of this figure is available in the online issue.)