Fig 2: Interaction between 14-3-3ε and CD13. (A) Representative SPR sensorgrams showing the binding of recombinant human 14-3-3ε (red, 10 µM), human anti-CD13 antibody SJ1D1 (blue, 1 µg/ml), and isotype IgG1 control (green, 1 µg/ml) on recombinant human CD13 immobilized on the sensor chip. The differential response was obtained by subtracting the signal in the blank channel from that in the experimental channel. Response is expressed in resonance units (RU) relative to time in seconds. (B) Kinetics of 14-3-3ε binding to CD13. Six concentrations of 14-3-3ε (5, 7.5, 10, 15 and 20 µM) were tested. The base line (in red) corresponds to HBS-EP alone. Software analysis gave a KD value of 2.45×10−6 M with χ-squared value 2.09.

Fig 3: Different expressions of miR-483-5p in the mouse model of diabetes in the exosomes derived from renal tissues, urine, and renal tubular.A Detection of CD63 and APN protein levels by western blot. B Detection of miR-483-5p expression by qRT-PCR. **P < 0.01 vs. control or db/m. Student’s t-test. of three independent experiments. STZ Streptozotocin, db/m db/misty, APN Aminopeptidase N.

Fig 4: HPL therapy reverses OVX-induced endometabolite imbalances in the glutathione metabolism pathway. (A) PLS analysis of the metabolomic profiles of urine and serum samples; the differences among control (blue dot), OVX (red square), HPH (HPL-H, green triangle), and HPL (HPL-L, yellow downward triangle) samples are shown. Each symbol on the 2D score plot represents a result for an individual. (B) Gene-metabolite KEGG pathway enrichment plot of the biomarkers in each pathway; the bar length (blue: enrichment and orange: topology) represents the significance levels of the pathways. (C) Redox GSH/GSSG ratio and relative peptide quantification for Ggt1 and Anpep. The data are representative of three experiments. The data are pooled from three experiments (n = 3, mean ± s. e.m.). (D) and (E) Quantification of metabolites from glutathione metabolism in 3, 6, and 12 weeks urine samples and 12 weeks serum samples using a liquid chromatography–tandem mass spectrometry analysis. The data were pooled from three experiments (n = 9, means ± s.d.). HPL therapy dosage: HP30, 30 mg/kg/d; HP10, 10 mg/kg/d. *p < 0.05, **p < 0.005, and ***p < 0.0005 compared with control rats; #p < 0.05, ##p < 0.005, and ###p < 0.0005 compared with OVX rats. NS, not significant.

Fig 5: 14-3-3ε binding to the chondrocyte surface. Mouse chondrocytes were treated with biotin–14-3-3ε for 15 min in the absence (A) or in presence of excess 14-3-3ε (B) or anti-CD13 blocking antibody (AB108310; Abcam) (C). Cells not treated with biotin–14-3-3ε were used as a negative control (D). Cells were analyzed by immunofluorescence microscopy, with Alexa-Fluor-568-conjugated streptavidin or Alexa-Fluor-488-conjugated agglutinin as the plasma membrane marker.