Figure S6.

Characterization of mitochondrial dynamics, insulin, and noradrenergic responses upon differentiation of BA with T₃, insulin, and rosiglitazone-enriched medium. (A) TOM 22 (left) and COX-IV immunofluorescence (right) in BA differentiated with medium 1 or 2 (day 7). Graph shows MitoTracker Green labeling by flow cytometry (n = 4 independent experiments). (B) p-DRP1 (pink) and MFN2 (green) immunofluorescence, western blotting, and quantification (n = 3–4 independent experiments). Nuclei staining with DAPI (blue). Scale bars represent 20 μm. (C) Western blotting and quantification showing the fold increase of IR and AKT phosphorylation after insulin stimulation (n = 3 independent experiments). (D) Western blotting and quantification of PTP1B (n = 4 independent experiments). (E) (left) Ucp1 mRNA (n = 3 independent experiments) and UCP-1 protein levels (n = 5 independent experiments) after stimulation of BA with NE after differentiation with medium 2. (right) Glycerol released by BA differentiated with medium 2 after stimulation with NE (n = 3 independent experiments). (F) NE-induced Ucp1 mRNA levels in BA differentiated with medium 2vs medium 1 (n = 3 independent experiments). Results are expressed as mean ± SEM. Statistical analysis in A-B was performed by two-way ANOVA. ^∗Comparison between same genotype and different media. ^#Comparison between genotypes differentiated with the same medium. The statistical analysis in C–F was conducted with the Mann–Whitney U test. ^∗Comparison between genotypes. ^∗,#p < 0.05, ^∗∗,##p < 0.01, and ^{∗∗∗,###}p < 0.001.