Figure 3.

SIRT1 overexpression conferred protection against insulin resistance, but not UCP-1 induction, in brown adipocytes under pro-inflammatory conditions. (A) Western blotting and graphs showing IR and AKT phosphorylation in BA with or without insulin (n = 3–4 independent experiments). (B) Western blotting and quantification of PTP1B and SIRT1 (n = 7–8 independent experiments). (C) Western blotting and quantification of fold change of UCP-1 (protein and mRNA) after NE stimulation (n = 3–4 independent experiments). (D) Glycerol released after NE stimulation (n = 3 independent experiments). (E) Western blotting and quantification of insulin-induced IR and AKT phosphorylation after CM-CTR or CM-LPS treatment (n = 3–4 independent experiments). (F) The left graph represents the basal ³H-2-deoxyglucose uptake in BA (n = 4 independent experiments) and the right graphs represent the fold increase in insulin-induced glucose uptake after treatment with CM-CTR or CM-LPS for 18 h and further stimulated with insulin (10 nM) (n = 4 independent experiments). (G) Western blotting and graphs showing the fold increase in UCP-1 (protein and mRNA) after NE stimulation (n = 3–5 independent experiments). Results are expressed as mean ± SEM. The statistical analysis in A, E, F (left graph), and G (right graphs) was performed by two-way ANOVA. ^∗Comparisons between treated and untreated BA with the same genotype. ^#Comparison between genotypes receiving the same treatment. Statistical analysis in B-D was conducted with Mann–Whitney U test. ^∗Comparison between genotypes. Statistical analysis in F (right graph) and G (left graph) was performed with the Mann–Whitney U test. ∗Comparison between treatments. ^∗,#p < 0.05, ^∗∗,##p < 0.01, and ^{∗∗∗,###}p < 0.001.