STING depletion promotes AMPK and its substrates activation in mice muscle. (A–F) Representative Western blots (n = 3; A) and quantification (n = 6–8) of the phosphorylation of AMPK (at Thr172; B) and its substrates TBC1D1 (at Ser237; C), Raptor (at Ser792; D), TSC2 (at Ser1387; E), and ACC (at Ser79; F) in gastrocnemius muscles from indicated genotypes before and after exercise. Data are represented as mean ± SEM. ST, STING; NE, non-exercise; EX, exercise; NS, not significant. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Unpaired two-tailed t test for comparison of two groups. Two-way ANOVA for comparison of magnitude of changes between different groups in mice of different genotypes. ^#, P ≤ 0.05; ^###, P ≤ 0.001. (G) Increased GLUT4 translocation to plasma membrane in STING KO mice after exercise. Representative images of GLUT4 co-localization of Laminin2 in tibialis anterior muscles (n = 4). Scale bar, 20 μm. (H) Quantification of GLUT4 co-localization of laminin2 in tibialis anterior muscles (n = 4; G) are represented as mean ± SEM. An area of 220.5 mm² per mouse was evaluated. NS, not significant; **, P ≤ 0.01. Unpaired two-tailed t test for comparison of two groups. (I) AMPK activation analysis in STING WT, STING KO, and STING KO–STX17 KO cells. The STING WT, STING KO, and STING/STX17 DKO MEF cells were incubated in full medium or HBSS for the indicated time. AMPK phosphorylation (at Thr172) was detected by Western blot. Representative images were shown. The experiments were repeated three times and similar results were observed. Normalized fold change of p-AMPK/AMPK was listed. Source data are available for this figure: SourceData F4.