Fig 2: The combination treatment of Arazyme (Ara) and ESLs enhanced hepatic β-oxidation. (A) Hepatic mRNA expression levels detected by real-time qRT-PCR and normalized using Gapdh as a reference gene. (B) Hepatic protein expression levels detected by western blotting with anti-PPARα, anti-UCP2, and anti-GAPDH antibodies. Values are presented as means ± SE (n = 6). * p < 0.05, ** p < 0.01 vs. HFD group.

Fig 3: Regulation of UCP2 is AMPK- and PPARγ-dependent in MTO1 fibroblasts. (A) qRT-PCR analysis of the UCP2 mRNA expression in MTO1 human fibroblasts (MTO1 HF), treated or not with 5 μM rosiglitazone (RGZ) for 1 h. Data are represented as fold change respect to WT HF values. (B) Representative immunoblots of phosphor-Thr172-AMPKα, AMPKα and UCP2 in WT HF and MTO1 HF, treated or not with 5 μM RGZ for 1 h. Porin and AMPKα were used as loading controls. Full-length western blots are included in supplementary information (Fig. S23). The scatter plot shows densitometric data for UCP2 normalized to porin and phosphor-Thr172-AMPKα normalized to AMPKα, and represented as fold change relative to WT HF. (C) Representative immunoblots of phosphor-Thr172-AMPKα, AMPKα and UCP2 in WT HF and MTO1 HF, treated or not with 1 mM AICAR for 1 h. Porin was used as a loading control. Full-length western blots are included in supplementary information (Fig. S23). The scatter plot shows densitometric data for UCP2 normalized to porin and phosphor-Thr172-AMPKα normalized to AMPKα, and represented as fold change relative to WT HF. All data are the mean ± SD of at least three different experiments. Differences from WT HF values were found to be statistically significant at *p < 0.05, **p < 0.01 and ***p < 0.001.

Fig 4: Effects of extracts of chickpeas (ECP) on carnitine palmitoyltransferase 1 (CPT1) and UCP2. Western blot analysis was performed for (A) CPT1 and (C) UCP2. (B and D) Densitometric analysis was performed by reading individual protein bands thereafter. Each value was corrected with the internal standard RPS18, and the value relative to day 0 was then calculated (means ± SD of triplicate experiments; Jonckheere-Terpstra test: CPT1, *p=0.0124; UCP2, *p= 0.0369).

Fig 5: Activation of UCP2 by ROS is required for endogenous H2S to facilitate microglial phagocytosis of RBCs and spontaneous hematoma resolution following ICH. (A and B) Representative fluorescence images and quantification data showing that the RBC lysate-induced reduction in ΔΨm in primary microglia was blocked by genipin, an inhibitor of ROS-mediated activation of UCP2 (JC-1 staining, n = 9 from 3 independent biological replicates). (C and D) Genipin inhibited the engulfment of RBCs by primary microglia treated with RBC lysate, as indicated by the number of nonphagocytosed RBCs and the phagocytosis index (n = 3). (E and F) Quantification data and representative fluorescence images showing that knockdown of UCP2 in microglia blocked the reduction in ΔΨm induced by RBC lysate in primary microglia (TMRM staining, n = 9 from 3 independent replicates). (G) Immunoblot analysis of UCP2 expression in the striatum transfected with a lentivirus expressing UCP2-shRNA or NC-shRNA for 14 days (representative images from 3 independent experiments). Ipsil: the striatum ipsilateral to lentiviral injection. Contra: the contralateral striatum. (H and I) Striatal knockdown of UCP2 delayed spontaneous hematoma resolution following ICH, and the delay was not rescued by the H2S donor ADT, as indicated by hemorrhagic volumes at days 5 and 14 following ICH (n = 6).