Fig 4. ACLP expression increases in the stromal vascular fraction from diet induced fibrotic epididymal adipose tissue.

A, epididymal adipose tissue from C57BL6/J male mice on low fat diet or high fat diet for 16 weeks (n = 3 for each diet group) was excised and fixed with methyl Carnoy, stained with picrosirius red and imaged using brightfield (left panels) and under polarized light (right panels). B, Tissue sections of epididymal adipose tissue following low fat diet or high fat diet were immunostained for ACLP and perilipin and counterstained with DAPI. Data are representative of parallel 10 μm sections. Closed arrowhead indicates vasculature. Open arrowhead indicates peri-cellular staining. Scale bar represents 100 μm. C, Epididymal adipose tissue from C57BL6/J male mice on low fat diet or high fat diet for 16 weeks (n = 3 for each diet group) was excised, enzymatically digested, fractionated based on buoyancy and then protein lysates were generated of each population. Whole stromal vascular fraction lysate was analyzed by SDS-PAGE and Western blot with antibodies against ACLP, α-SMA, desmin, SM22, CRP2 and CypA. Protein expression was quantified by densitometry normalized to CypA expression and relatively compared to higher expressing diet sample. *, p < 0.05 versus low fat diet by Students t-test. D, Whole adipocyte lysate was analyzed by SDS-PAGE and Western blot with antibodies against adiponectin, PPARγ and CypA. Protein expression was quantified by densitometry normalized to CypA expression and relatively compared to low fat diet. *, p < 0.05 versus low fat diet using Students t-test.