D12492

C57BL/6J, CCR2^−/−, and CX₃CR‐1^GFP (B6.129P2(Cg)‐Cx3cr1^tm1Litt/J) mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA). C57BL/6 CD169‐DTR mice were generated in our laboratory (Purnama et al, 2014 (link)) and crossed with CCR2^−/− to obtain the final CCR2^−/−CD169‐DTR mouse line. We used male mice only for these experiments. To induce obesity, mice aged 5–6 weeks were given a high‐fat diet (HFD) that comprised 60% kcal of fat (Research Diet, D12492, New Brunswick, NJ, USA) for 8, 12, or 16 weeks; age‐matched control mice were given a normal chow diet (ND).
CD169‐DTR⁺ and CD169‐DTR⁻ (WT) mice were intraperitoneally (i.p.) injected with 20 ng/g DT (Sigma) every 3–4 days to maintain the depletion of CD169⁺ ATMs. C57BL/6J mice were i.p. injected with rat IgG2a isotype control (BioLegend) or anti‐CSF1R blocking antibody (Clone AFS98, BioXCell, West Lebanon, NH, USA), twice per week (400 μg per mouse) for 12 days. No randomization nor blinding was used during our experiments.
All mice were bred and maintained in a specific‐pathogen‐free animal facility at Nanyang Technological University (NTU), Singapore. The mice experiments were carried out in strict accordance with the recommendations of the NACLAR (National Advisory Committee for Laboratory Animal Research) and approved by the Institutional Animal Care and Use Committee (IACUC) of NTU (ARF‐ SBS/NIE A18014).