Fig 2: GLUT4 is localized to the plasma membrane in Arid5b-/- GC muscles. a Coimmunofluorescence analysis was performed for GLUT4 (green) and dystrophin (red) in GC muscle sections, and representative confocal images are shown. Merged images show increased colocalization of GLUT4 and dystrophin at the plasma membrane in Arid5b-/- GC muscles relative to Arid5b+/+ GC muscles. b The Pearson’s correlation coefficient quantifying the degree of colocalization was calculated (n > 10 images per mouse, n = 3 mice per genotype). Data are presented as the means ± SD. *, P < 0.05

Fig 3: D-Chiro-Ins does not modulate the expression of GLUT4. Mature SGBS adipocytes cells were treated with D-Chiro-Ins 10-8 M, E2 10-9 M or INS 10-7 M, and combinations for 24 h. Cell lysates were immunoblotted with an anti-GLUT4 antibody, anti-p-GLUT4 antibody, and anti-GAPDH antibody. (A) Panels show representative blot of GLUT4, p-GLUT4 S488 and GAPDH. (B–D) Analysis of the relative expression level of GLUT4, p-GLUT4 and the ratio p-GLUT4/GLUT4; expression that was normalized with GAPDH and relative to the control. Values are presented as mean ± SEM of three independent experiments. Results were analyzed statistically by one-way RM ANOVA followed by Dunnett’s multiple comparisons test (*p < 0.05, **p < 0.001 versus control). D-Chiro-Ins, D-chiro-Inositol; E2, 17ß-estradiol, INS, insulin; GLUT4, Glucose Transporter 4; p-GLUT4, phospho-GLUT4; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Fig 4: Downregulation of TBC1D1 expression in Arid5b-/- GC muscles. The expression of TBC1D1 protein (a), TBC1D4 protein (b) is shown. Protein expression was normalized to the corresponding loading control (HSP90, HSP70 or vinculin), and the data are presented as the means ± SD (n = 6). ***, P < 0.001. c, d The expression of GLUT4 and GLUT1 protein in GC muscles is shown. Quantification of GLUT4 and GLUT1 protein in GC muscles is shown. (bottom) Quantitation of GLUT4 protein expression was performed, and the data are presented as the means ± SD (n = 6)