Figure 1. Inducible PTEN elimination from the adipocyte.

(a) Quantitative PCR results on a floated adipocyte fraction showing PTEN gene expression from mice fed a doxycycline (600 mg kg⁻¹) diet for 4 or 30 days (mean of N=2 for each bar). (b) Gonadal adipose tissue immunofluorescence using a PTEN antibody (Abcam 32199) with 4,6-diamidino-2-phenylindole (DAPI) counterstain to show nuclei. (Scale bar, 152 μm for × 4, 65 μm for × 15.) (c) Phospho-AKT, total AKT and actin for isolated adipocytes stimulated with insulin (1 nM) for 10 min (d) Western blot of protein extracts from gonadal adipose tissue harvested from mice stimulated with insulin for the indicated amount of time. Staining was then performed against phospho-AKT (S473), total AKT, p44/42 or total actin as indicated and then quantified using a Licor Imager (1 mouse for each group (control or AiPKO) at each time point, 0, 5, 10 or 15 min). (e) Western blot for plasma adiponectin levels from mice after 4 weeks of a doxycycline-containing chow diet (N=3 for each group). (f) Blood glucose of mice fasted for 4 h (N=4 for each group). (g) Oral glucose tolerance test of mice on a doxycycline-containing chow diet for 4 weeks (average area under the curve—control=28,588, AiPKO=17,001, P value=0.0221; Student's T-test) (N=3 for each group). All data compare control mice with AiPKO mice. For all points, *P<0.05, **P<0.01 (Student's T-test). All data are mean±s.e.m.