Journal: medRxiv
Article Title: Physiological and molecular characterization of individuals carrying a diabetogenic mtDNA mutation establishes a mitochondrial basis for insulin resistance in humans
doi: 10.64898/2025.12.17.25342274
Figure Lengend Snippet: (A) Schematic overview of the canonical signaling events modulating muscle insulin action and its interaction with mTORC1 signaling. (B to D) Canonical insulin signaling as determined by phosphorylation of Akt2 on Ser473 and Thr308, and phosphorylation of the Akt substrates GSK3β on Ser9 and TBC1D4 on Thr 642 in whole-muscle lysates from skeletal muscle biopsy samples obtained before (Basal) and immediately after (Insulin) the hyperinsulinemic-euglycemic (HE) clamp. Levels of total Akt2, GSK3β, and TBC1D4 were comparable across groups and unaffected by insulin. (E and F) mTORC1 signaling as determined by phosphorylation of p70 S6 kinase (p70S6K) on Thr389 and S6 on Ser235/236 in whole-muscle lysates from skeletal muscle biopsy samples obtained before (Basal) and immediately after (Insulin) the HE clamp. Levels of total p70S6K and S6 were comparable across groups and unaffected by insulin. Linear mixed models were used to estimate within– and between-group differences. Data are presented as observed individual values (with lines connecting individually matched participants) and estimated means ± 95% confidence limits, unless otherwise stated. For m.3243A>G carriers, individual datapoints are color-shaded to indicate muscle mtDNA heteroplasmy (light red = low, dark red = high). *Different from Basal ( P < 0.05). Basal, n = 30; Insulin, n = 27 (13 in m.3243A>G, 14 in Controls). Illustrations in (A) were created with BioRender.com.
Article Snippet: The following antibodies were used: Phospho-Akt (Thr308) (Cell Signaling, catalog no. 9275); Phospho-Akt (Ser473) (Cell Signaling, catalog no. 9271); Akt2 (Cell Signaling, catalog no. 3063); Phospho-GSK-3β (Ser9) (Cell Signaling, catalog no. 5558); GSK-3β (BD Transduction Laboratories, catalog no. 610202); Phospho-TBC1D4 (Thr642) (Cell Signaling, catalog no. 8881); TBC1D4 (Abcam, catalog no. ab189890); Phospho-p70S6K (Thr389) (Cell Signaling, catalog no. 9205); p70S6K (Cell Signaling, catalog no. 9202); Phospho-S6 (Ser235/236) (Cell Signaling, catalog no. 4858); S6 (Cell Signaling, catalog no. 2217); PRDX2 (Abcam, catalog no. ab109367); PRDX3 (Abcam, catalog no. ab73349).
Techniques: Phospho-proteomics
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