Fig 2: Western blot detection and quantification of ERK1/2 (A‐B) and p38 mitogen‐activated kinase (C‐D) phosphorylation in brown preadipocytes treated with NPB (100 nm) for the indicated time points. Effect of SB239063 blocker (E) on Ucp1 mRNA expression in cells differentiated with or without NPB (100 nm) for 1 day. Results are shown as the mean ± SEM [n = 3 (western blot), n = 6 (mRNA expression)]. Statistical differences were determined by one‐way ANOVA followed by the Bonferroni post hoc test (B‐E). *P < 0.05 or **P < 0.01 vs vehicle (water) treated cells.

Fig 3: Renal ischemia reperfusion (IR) induces downregulation of UCP1 in mice. (A) A volcano plot showed the differentially expressed mRNAs in IR group compared to Sham group. Red and green dots represent upregulated and downregulated mRNAs in kidneys 24 h after ischemia reperfusion (IR), respectively (fold change =2.0 and P-value =0.05). Black dots represent the genes that were not differentially expressed. UCP1 was the most downregulated gene. (B) Quantitative RT-PCR analysis of the top 11 downregulated mRNAs in the mouse kidneys 24 h after renal IR. (C) Immunohistochemical staining for UCP1 in multiple organs in mice. UCP1 was specificly expressed in kidney, including renal cortex and medulla (arrow). Scale bar, 100 µm. (D) Quantitative RT-PCR analysis of UCP1 mRNA during renal IR. (E) Enzyme-linked immunosorbent assay (ELISA) of UCP1 in kidney. Data represent mean ± SEM. n = 4–6. **P < 0.01 versus Sham group. (F) Representative images of immunohistochemical staining for UCP1 in kidneys from patients with acute kidney injury (AKI) or normal controls. UCP1 was expressed in renal tubular epithelium (arrow). Scale bar, 100 µm. The control samples were from normal portions of nephrectomy specimens that had been removed for localized renal tumor. Acute tubular necrosis (ATN) was confirmed by renal biopsy in the patients with AKI.

Fig 4: Deletion of UCP1 aggravates acute kidney injury and increased reactive oxygen species generation. (A) Serum creatinine 24 h after renal IR. 25-min renal ischemia induced a significant increase in serum creatinine in UCP1-/- mice, but not in wild-type (WT) mice. (B) Serum creatinine 72 h after cisplatin injection. Treatment with 20 mg/kg cisplatin (Cis20) caused a significant increase in serum creatinine in UCP1-/- mice, but not in WT mice. (C) Malondialdehyde (MDA) concentration in mice kidneys. (D) Images and quantification of dihydroethidium (DHE) staining for detection of reactive oxygen species (ROS) generation in mice kidneys. Scale bar, 100 µm. Data represent mean ± SEM. n = 6. *P < 0.05, **P < 0.01 versus WT-IR25’ or WT-Cis20’ group.

Fig 5: The effects of 4 weeks of resistance training on protein expression in epididymal white adipose tissue (eWAT). Representative Western blot images of each specific band (a). Protein contents of OXPHOS (b), PGC‐1α, TH, and SERCA2 (c) in eWAT. Ponceau staining was used as a loading control. UCP1 mRNA expression in eWAT (D). n = 6–8 in each group. Mean ± SEM