Clone fa 11

Aliquots of epididymal and subcutaneous adipose were fixed in 4% PFA then washed in PBS. Adipose was blocked and permeabilized in 5% BSA, 0.3% Triton in PBS before incubating overnight with CD68-PE conjugate (Biolegend, Clone FA-11) and Isolectin GS-IB₄ AF488 conjugate (Thermofisher) or CD31 AF488 (Biolegend, Clone MEC13.3) at 4°C. After a final wash, samples were mounted in a 1:1 solution of PBS: Glycerol and digital images were acquired using confocal microscopy (Nikon Instruments Incorporated, Model TE200-E2; 20X objective). Images were processed using ImageJ software.

Aliquots of epididymal and subcutaneous adipose were fixed in 4% PFA then washed in PBS. Adipose was blocked and permeabilized in 5% BSA, 0.3% Triton in PBS before incubating overnight with an anti-CD68-PE conjugated antibody (Biolegend, Clone FA-11) and an Isolectin GS-IB₄-AF488 conjugate (Thermofisher) at 4°C. After a final wash, samples were mounted in a 1:1 solution of PBS:Glycerol and digital images were acquired using confocal microscopy (Nikon Instruments Incorporated, Model TE200-E2; 20X objective). 40 μm Z-stacks with 2 μm step size were acquired with a 20x magnification power and processed using ImageJ software.