Figure 5

Tracking labeled adiponectin oligomer circulatory clearance and tissue uptake over time. A) Representative IR800-labeled adiponectin (green) and IR700-labeled IgG (red) over time from sera separated by gel electrophoresis; Iso: isosorbide dinitrate treatment. B) Three hours after injection, and following PBS perfusion, whole tissue was scanned to assess potential changes in adiponectin (green) localization and tissue homogenates, normalized for total protein concentration, were measured by dot blot quantification. Data are presented as adiponectin:IgG (red) ratio. C) Adiponectin:IgG ratios in sera over time following HMW (red lines) adiponectin or LMW (green lines) injection following acute vehicle (solid lines) and isosorbide dinitrate (Iso; dotted lines) administration. D) The half-life of HMW adiponectin was significantly reduced with iso treatment. E) Adiponectin:IgG ratios in tissues 3 hours following HMW adiponectin injection and normalized to liver values show no differences. F) Similarly, no differences were noted for LMW tissue uptake with isosorbide dinitrate. G) Adiponectin:IgG ratios in sera over time following HMW (red lines) adiponectin or LMW (green lines) injection into chow fed and PPARγ agonist (nTZD) diet fed (dotted lines) mice showed a delay in HMW clearance. H) Quantified half-lives demonstrated a significant decrease in HMW adiponectin clearance from circulation. I) nTZD treatment increased HMW adiponectin uptake in mouse brains. J) Conversely, nTZD-treated brains had significantly less LMW adiponectin uptake. Quadriceps also contained less adiponectin with the PPARγ agonist. Half-lives quantified by non-linear regression (decay) analysis. All values compared by two-tailed student’s t-test with unequal variance. *p < 0.05 between treatment and control.