Figure 6. Increased Glucose Uptake in P-BCA Muscles with Activation of AMPK, but Not Wnt signaling.
(A) Fed GTT of control and P-BCA mutant mice. n ≥ 8 per cohort, repeated ≥ 3 cohorts.
(B, C, D) Hyperinsulinemic-euglycemic clamps of control and P-BCA siblings. Insulin and glucose infusions were started at time 0, 5 hours after food removal. (B) Blood glucose levels and (C) glucose infusion rate (GIR) 15 minutes before and during clamp. (D) Glucose uptake of indicated muscles (Vastus = vastus medialis, Gastroc = gastrocnemius) derived from isotope incorporation 30 minutes after injection of 14C 2-deoxyglucose at time 120 minutes of clamp. n ≥ 10, repeated ≥ 2 cohorts.
(E) Western blots of proteins isolated from fed and fasted (4 hours) sibling 4-month old male gastrocnemius muscles probed with the indicated antibodies. C = control, M = P-BCA mutant, p = phosphorylated, t = total.
(F) AMPK blots of gastrocnemius muscles of P-BCA siblings injected with vehicle (P-BCAVeh) or Compound C (P-BCA-CC), p = phosphorylated, t = total.
(G) Fasting blood glucose levels of P-BCA males injected with vehicle (P-BCA-VH) or Compound C (P-BCA-CC). Mice were injected with vehicle or Compound C at time 0 then fasted for three hours.
(H) GTT of P-BCA males injected with vehicle (P-BCA-VH) or Compound C (P-BCA-CC) and fasted for four hours.
(I) Fractionation of cytoplasm and plasma membrane from limb muscles of control (C1-C4) and P-BCA (M1-M4) male siblings both with and without STZ. Cadherin and the insulin receptor (IR) are plasma membrane markers, tubulin is a cytoplasm marker.
(J) IGW and muscles of control (CON) and P-BCA mice were removed and incubated in X-gal. Blue color reflects lacZ expression.
(K) Western blot of muscle extracts of 4 control and 4 P-BCA siblings probed with anti-β-catenin and anti-tubulin antibodies. β-catenin indicates wild-type form; BCA indicates activated form. Tubulin serves as loading control. P-BCA adipose depot extracts are BCA positive control.
(L) qPCR of control and P-BCA muscle extracts for the indicated β-catenin targets. n ≥ 8 per cohort, repeated ≥ 3 cohorts.
Error bars indicate SEM. Statistical significance assessed by two-tailed Student’s t-test, *p<0.05, **p<0.01.