Fig 1: Effect of JTXK granules on AMPK/SIRT1/PGC-1α signaling in skeletal muscle of HFD-induced diabetic mice. The relative protein expression levels of AMPKα, SIRT1, PGC-1α, CPT1, UCP3, and PPARα (both in cytosolic and in nuclear) in the skeletal muscle (a–h) and GLUT4 membrane translocation (i) were determined by western blotting. Gene expressions of AMPKα, CPT1, PGC-1α, PPARα, SIRT1, and UCP3 (j–o) were determined by RT-PCR. NC, DM, Met, and JTXK refer to the normal control, diabetes, metformin, and Jiang Tang Xiao Ke granule groups, respectively. Data were presented as mean ± SD. n = 6 in each group. ∗P < 0.05, compared with the NC group. #P < 0.05, compared with the DM group.
Fig 2: Immunofluorescence analysis in VAT and representative photomicrographs at magnification ×10 of: (A) ADRB1, (B) UCP2 and (C) UCP3. The counterstaining of nuclei (Hoescht) is shown in blue and PLIN1 in red. Quantifications are presented as the average fluorescence intensity ± SD and compared by Student’s t-test. * = p < 0.05, ** = p < 0.001.
Fig 3: Effect of JTXK granule drug-containing serum on AMPK/SIRT1/PGC-1α signaling in IR C2C12 cells. (a–g) Protein expressions of AMPKα, SIRT1, PGC-1α, PPARα, UCP3, and CPT1 measured by western blotting. (h–m) Relative gene expressions of AMPKα, SIRT1, PGC-1α, PPARα, UCP3, and CPT1 measured by RT-PCR. Con, Met, and DCS mean the control, metformin, and drug-containing serum of Jiang Tang Xiao Ke granules, respectively. Data were presented as mean ± SD. ∗P < 0.05, compared with the Con group.
Fig 4: UCP3 expression, mitochondrial membrane polarization, and metabolic profile of human spermatozoa. (A) UCP3 relative expression in spermatozoa from normozoospermic versus asthenozoospermic men. (B) Evaluation of mitochondrial membrane potential using the JC-1 dye after inhibition of UCPs by increasing concentrations of genipin (0.5, 5, and 50 µM) for 3 h. (C) Lactate and (D) acetate production by human spermatozoa after UCPs inhibition by increasing concentrations of genipin (0.5, 5, and 50 µM) for 3 h. Results are expressed as Tukey’s boxplot (median, 25th to 75th percentiles ± 1.5 IQR). +−Represents group average. Significantly different results are expressed as: *−Relative to the control group.
Fig 5: Immunofluorescence staining for UCP1, UCP2, and UCP3 protein expression in human spermatozoa. (A) Negative control. Immune labeling of UCP1 (B), UCP2 (C), and UCP3 (D) was observed in human spermatozoa. Scale bar—10 µm.
Supplier Page from Abcam for Anti-UCP3 antibody