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. 2015 May 6;64(9):3081–3092. doi: 10.2337/db14-1810

Figure 4.

Figure 4

Insulin signaling and GLUT4 translocation are not affected in skeletal muscle of SIRT3 KO mice. A: Gastrocnemius protein homogenates obtained from 5-h–fasted (Basal) or clamped (Insulin) HF-fed mice were applied to a 4–12% SDS-PAGE (n = 4). Western blotting was performed for Akt, P-Akt (Ser473), AS160, and P-AS160 (Thr588). Integrated intensities were obtained by the Odyssey software. B: Immunoprecipitation (IP) from gastrocnemius muscle extracts obtained from clamped (Insulin) mice were performed with total IRS1 antibody; then, immunoblots were completed for IRS1, P-IRS1 (Ser612), and the p85 subunit of PI3K (n = 6). Integrated intensities were obtained by the ImageJ software. C: Western blotting was performed for GLUT1 and GLUT4 on gastrocnemius protein homogenates obtained from 5-h–fasted (Basal) or clamped (Insulin) mice (n = 4). Integrated intensities were obtained by the Odyssey software and normalized to GAPDH intensities. D: Confocal imaging of GLUT4 and plasma membrane marker caveolin-3 (CAV3) was performed on clamped (insulin) gastrocnemius cryosections (n = 5–8). *P < 0.05.