Fig 2: Activation of the RAS/ROS and impaired Glut4 trafficking in the skeletal muscle of 5/6Nx rats with high-salt intake are prevented by blockade of a renal/adipose-cerebral-muscle sympathetic reflex.(a and b) Overexpression of skeletal muscle (SM) AGT (a) and Ang II (b) in 5/6Nx rats under high-salt condition was inhibited by intracerebroventricular administration (ICV) of losartan or clonidine, denervation of the kidneys (RDX) or epididymal fat pads (Epi DN), or deafferentation of epididymal fat pads with resiniferatoxin (Epi RTX). (c–e) Norepinephrine (NE) level(c), and expression of TNFa (d) and Nox2 (e) in SM. (f) Glut4 expression in extracted plasma membrane (PM) and the ratio of PM to total membrane (TM) Glut4 under insulin stimulation. (g) Hindlimb perfusion measured by contrast-enhanced ultrasound under insulin stimulation. (h) Insulin-stimulated 2-deoxy-glucose (DG) uptake in SM. (i) Steady-state glucose infusion rate (GIR) during hyperinsulinemic-euglycemic clamp. Data are expressed as mean ± SD (n = 6 in each group). *P < 0.05 vs 5/6Nx rats under high-salt condition given vehicle (0 mg/kg/d inhibitor) by unpaired t-test.

Fig 3: Changed protein expression in cardiac muscles by 10% glucose administration for heart failure deteriorated by high-dose liraglutide. a, b GLUT-4 and CD36 were upregulated further. c, d The levels of the apoptotic marker cleaved caspase-3 and autophagy marker LC3 II/I significantly reduced with 10% glucose administration. n = 6 in each group

Fig 4: Aerobic exercise attenuated insulin resistance through the upregulation of IRS--1/PI3K/AKT signaling. (A) Western blot images and quantifications of muscle samples, including: (B) IRS--1, (C) PI3K, (D) p--AKT/AKT, and (E) GLUT4. (F) mRNA expression of GLUT4 in skeletal muscle. Values are expressed as the mean ± SD. CON, normal control; HFD, high-fat diet; S, sedentary; E, exercise. ** p < 0.01 vs. NS group; # p < 0.05, ## p < 0.01 HS group (n = 6).

Fig 5: Gq-meditated stimulation of AMPK phosphorylation, GLUT4 translocation, and phosphorylation of HSL (S565) in 3T3F442A mouse adipocytes.a Western blotting studies with GqD-expressing mouse 3T3F442A cells (GqD-3T3F442A cells) treated with CNO (10 µM) for different periods of time. b Quantitative analysis of pAMPK/AMPK and pAS160/AS160 protein expression levels shown in (a). Data shown in (b) were derived from four of five independent experiments. Protein expression levels were normalized relative to protein expression levels at time ‘0’. c Western blotting studies carried out in the presence of a Gq/11 inhibitor. GqD-3T3F442A cells) were incubated with CNO (10 µM) for 30 min either in the absence or presence of FR900359 (FR; 1 µM), a Gq/11 inhibitor. d Quantitative analysis of pAMPK/AMPK and pAS160/AS160 protein expression levels shown in (c). e Immunoblotting analysis of GLUT4 plasma membrane localization. After treatment of GqD-3T3F442A cells with CNO (10 µM) or insulin (10 nM) for 30 min, the plasma membrane fraction was isolated and subjected to immunoblotting analysis using an anti-GLUT4 antibody. Na + /K + ATPase served as a marker for plasma membrane proteins. f Quantitative analysis of GLUT4 expression levels shown in (e). g Western blot analysis of HSL phosphorylation at position S565. GqD-3T3F442A cells were treated with CNO (10 µM), CL316,243 (CL; 100 nM), a ß3-adrenergic receptor agonist, or a combination of both drugs. To demonstrate the specificity of the HSL antibody used, GqD-3T3F442A cells were treated with HSL-specific siRNA or scrambled control siRNA. h Quantitative analysis of pHSL(S565)/HSL protein expression levels shown in (g). Data are presented as means ± s.e.m. b, d, f, h: one-way ANOVA followed by Bonferroni’s post-hoc test). Source data are provided as a Source data file.