Subcutaneous and perigonadal MMEKO preadipocytes show impaired insulin signaling. MMEKO and control (wild-type C57BL/6) mice aged 12 weeks were sacrificed for in vitro insulin signaling. The stromal vascular cells (preadipocytes) from the subcutaneous (inguinal) and visceral (perigonadal) adipose depots were isolated. Cells were serum-starved for 4 h before a 20-minute insulin stimulation followed by protein isolation and western blot. (A) Western blot of protein of subcutaneous and perigonadal MMEKO preadipocytes showed impaired response among different insulin-signaling proteins. (B–I) Densitometric analysis of protein levels. The levels of unphosphorylated proteins were grouped by adipose depot (PG = Perigondal, SC = Subcutaneous) and genotype (black = Control, grey = MMEKO). The levels of phosphorylated proteins were grouped by adipose depot, genotype, and insulin stimulation (– = no insulin, + = 20 min after 100 μM insulin). Asterisks indicate p ≤ 0.05 by Student's t-test. N = 3. Bars indicate mean ± s.e.m.