Fig. 3: Gq-meditated stimulation of AMPK phosphorylation, GLUT4 translocation, and phosphorylation of HSL (S565) in 3T3F442A mouse adipocytes.
From: Adipocyte Gq signaling is a regulator of glucose and lipid homeostasis in mice
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 β3-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.
