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Journal: iScience
Article Title: Functional and metabolomic analyses of brown adipose tissue during cold-deacclimation reveal rapid N-acetylated amino acid adaptations
doi: 10.1016/j.isci.2026.115146
Figure Lengend Snippet: Capacity for mitochondrial uncoupling and mitochondrial content regress in parallel during cold deacclimation (A) Schematic summary of the time points for sample collection. C57BL/6J mice were acclimated to the cold (4°C) for 7 days with ad libitum access to food and water and subsequently transferred to thermoneutrality (30°C) for 3 h, 12 h, 24 h, or 48 h. Diagram created with BioRender.com . (B) High-resolution respiratory flux per mg of saponin-permeabilized iBAT. Uncoupled respiration was measured in the presence of oligomycin and octanoylcarnitine. Complex I (CI) leak respiration was measured after the addition of malate-pyruvate-glutamate, and CI + CII leak and CI + II + G3P leak respiration were measured following the sequential additions of succinate and glycerol-3-phosphate for respectively, ( n = 16–18/group). (C) Immunoblotting of UCP1 protein expression ( n = 8/group). (D) Quantitative PCR (qPCR) of Ucp1 gene expression, ( n = 9/group). (E) BAT protein concentration (per mg tissue) was measured using a BCA assay in protein lysates, ( n = 10/group). (F) qPCR was used to determine the mtDNA:nDNA ratio (mt-ND1:n-HK2) ( n = 7/group). (G–J) TEM images were analyzed by quantitative morphometry for (G) mitochondrial surface area and (H) lipid droplet surface area, (J) Representative TEM images, ( n = 5 TEM images from 4 mice/group). Scale bars represent 2 μm. See also . Comparisons between time points were determined using a one-way ANOVA with Tukey post-hoc tests, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. All values are presented as means ± SD.
Article Snippet: Oxygen consumption was quantified in BAT using
Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Gene Expression, Protein Concentration, BIA-KA