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. 2021 Sep 25;297(5):101232. doi: 10.1016/j.jbc.2021.101232

Figure 5.

Figure 5

Macrophage depletion abolishes the exacerbated metabolic disorders in male GPNMB-KO mice fed an HFD.A, body weight of male WT and GPNMB-KO mice fed an HFD with or without treatment with clodronate liposomes (n = 10 for WT; n = 9 for KO; n = 9 for WT + Clodronate; n = 7 for KO + Clodronate). The body weight was similar between WT and KO mice in the same treatment group. Clodronate-treatment group showed significantly less body weight than in PBS group during 5 to 10 weeks of the HFD feeding. B, ITT in male WT and GPNMB-KO mice fed an HFD for 12 weeks with or without treatment with clodronate liposomes (n = 6 for WT; n = 7 for KO; n = 5 for WT + Clodronate; n = 5 for KO + Clodronate). C, IPGTT in male WT and GPNMB-KO mice fed an HFD for 12 weeks with or without treatment with clodronate liposomes (n = 8 for WT; n = 9 for KO; n = 7 for WT + Clodronate; n = 7 for KO + Clodronate). D, representative images of H-E staining and immunohistochemistry for F4/80 in the WAT of male WT and GPNMB-KO mice fed an HFD for 14 weeks with or without treatment with clodronate liposomes. Bars: 100 μm. E, quantitative analysis for F4/80-positive area in the immunohistochemistry shown in (D) (n = 6 each). F, representative images of immunohistochemistry for CD11c and F4/80 in the WAT of male WT and GPNMB-KO mice fed an HFD for 14 weeks with or without treatment with clodronate liposomes. Bars: 50 μm. G, quantitative analysis for CD11c- and F4/80-double positive or CD11c-negative and F4/80-positive areas in the immunohistochemistry shown in (F) (n = 5 each). H, quantitative PCR for TNF-α, IL-1α, IL-1β, IL-6, and CCL2 in the WAT of male WT and GPNMB-KO mice fed an HFD for 14 weeks with or without treatment with clodronate liposomes (n = 7 each). Data represent mean ± SDM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. #p < 0.05, ##p < 0.01, ###p < 0.001, and ####p < 0.0001. In B and C, ∗ for comparison between WT and KO; # for comparison between KO + PBS and KO + Clodronate. One-way ANOVA with Tukey’s post hoc test for multiple comparisons was used for the analysis of the differences between each group (E, G and H), while two-way ANOVA with post hoc analysis of Fisher’s LSD for multiple comparison was used for the analysis of the differences between each group (A–C). n.s., not significant.