Fig. 5: Thermogenesis by the Futile Creatine Cycle in classical brown adipocytes.
From: The Futile Creatine Cycle powers UCP1-independent thermogenesis in classical BAT
a Schematic of AAV-FLEX transduction and tamoxifen-inducible protein expression in iADKOCkb;Ucp1 mice. Created in BioRender. Kazak, L. (2025) https://BioRender.com/j99d203. b Representative oxygen consumption traces of 10,000 brown adipocytes from 10- to 12-week-old iADKOCkb;Ucp1 mice in response to noradrenaline (NA, 0.1 μM), oligomycin (Oligo, 15 μM), and SBI-425 (10 μM, added at the start). Bar graphs show basal, followed by acute additions of NA and oligomycin (n = 2 independent preparations/sex). c Basal-subtracted oxygen consumption rates from Fig. 5b. d Representative oxygen consumption traces of 10,000 brown adipocytes from 16-week-old wild-type male mice in response to NA, CCCP (40 μM), and rotenone (Rot, 5 μM). Bar graphs show basal, followed by subsequent stimulation with NA, CCCP, and Rot (n = 4). e Basal-subtracted oxygen consumption rates from Fig. 5d. f NA-stimulated glycerol release from brown adipocytes from 12-week-old iADKOCkb;Ucp1 mice (NA, 10 min) (n = 2 independent preparations/sex/group). Data calculated from Supplementary Fig. 6a. g Oxygen consumption rates of brown adipocytes from 16-week-old iADKOCkb;Ucp1 mice in response to NA and CCCP, grouped by treatments (L-GFP+veh, L-CKB+veh, L-GFP + SBI, L-CKB + SBI; n = 4, 6, 5, 7, respectively). h Basal-subtracted oxygen consumption rates from (g). i NA-stimulated glycerol release from brown adipocytes from 18-week-old iADKOCkb;Ucp1 mice (NA, SBI-425, 10 min) (n = 6: 2 females and 4 males for L-GFP groups; n = 8: 4 females and 4 males for L-CKB groups). Data calculated from Supplementary Fig. 6d. j in vivo NA-stimulated energy expenditure of 12-week-old iADKOCkb;Ucp1 mice expressing L-GFP or L-CKB in iBAT (females: n = 4 for L-GFP, n = 4 for L-CKB; males: n = 3 for L-GFP, n = 5 for L-CKB) acclimated to RT (23 °C ± 1 °C). b, c, g, h Two-way ANOVA (Tukey’s post-hoc test); f two-tailed student’s t tests; i two-way ANOVA (Fisher’s least significant difference (LSD)); j two-way ANOVA (Šidák post-hoc test). Data are presented as the mean ± s.e.m. and n numbers are of biologically independent experiments. Source data are provided as a Source Data file.
