Fig. 3

Sex- and age-related differences in the insulin signaling in gWAT. a–c Protein extract (100 μg protein) was separated by SDS-PAGE and subjected to Western blot analysis. We made tissue pools of 15 rats for p20 tissues and 5 rats for adult tissues. Results are product of three independent experiments. The bars represent the mean ± S.E.M. *P < 0.05, **P ≤ 0.005 compared to their control group by two-way ANOVA with post hoc Tukey's test; ^#P < 0.05, ^##P ≤ 0.007, ^####P < 0.0001 compared between sex and fasting vs fed state by two-way ANOVA with post hoc Tukey's test; d Adipocytes from gonadal depot were isolated with tissue digestion by collagenase P and incubated with insulin 1 nM or DMSO 0.1% for 30 min. Plasma membrane proteins were labeled with sulfo-NHS-SS-biotin. Labeled proteins (60 μg protein) were separated by SDS-PAGE and subjected to analysis by Western blot. Results are product of three independent experiments. The bars represent the mean ± S.E.M. ***P ≤ 0.0005, ****P < 0.0001 compared to their control group by two-way ANOVA with post hoc Tukey's test; ^#P < 0.05, ^##P < 0.005, ^####P < 0.0001 compared between sex and without stimulus vs with stimulus by two-way ANOVA with post hoc Tukey’s test; e Insulin-stimulated glucose uptake assay, gWAT (50 mg) was incubated for 30 min with 2-NBDG glucose 100 mg/mL (control condition) and with 2-NBDG glucose 100 mg/mL and insulin 100 nM (stimulating condition), n = 10 rats in each group. The bars represent the mean ± S.E.M. *P < 0.05, **P < 0.005 compared to their control group and compared between sex by two-way ANOVA with post hoc Tukey's test; f Summary of sex-related differences in the IR mechanism of PI3K/Akt signaling pathway in gWAT of p20 rats during postprandial state. Created with GraphPad Prism V 9.3.1 (471) and BioRender.com