differentiation 63 cd63  (Proteintech)

Journal: Journal of Sport and Health Science

Article Title: Long-term aerobic exercise enhances circulating exosomal miR-214-3p to promote endothelial progenitor cell-mediated repair of endothelial damage induced by obesity

doi: 10.1016/j.jshs.2025.101094

Figure Lengend Snippet: Eight weeks of aerobic exercise improved the proliferation and migration capabilities of circulating EPC in both humans and rats with obesity through circulating exosomes. (A) Representative transmission electron microscopy image of exosomes derived from human peripheral blood. Scale bar = 200 nm. (B) Exosome characterization and identification. Exosomes derived from human peripheral blood express TSG101 and CD63. (C) Nanoparticle tracking analysis confirms the presence of exosomes with a peak diameter of 100 nm, characteristic of exosomal size. Quantitative analysis of exosomes derived from human peripheral blood revealed no statistically significant difference in the number of exosomes isolated from equal volumes of circulating blood between the control group and the exercise group ( n = 30 for each group). (D) Cell proliferation assay results showed that exosomes derived from the exercise group significantly enhanced the proliferative capacity of human EPC compared to those from the control group, as measured by the CCK-8 method ( n = 20 for each group). *** p < 0.001, Exercise vs . Control. (E) Scratch assay results showed that exosomes derived from the exercise group significantly promoted the migratory ability of human EPC compared to those from the control group ( n = 5 for each group). * p < 0.05, Exercise vs . Control. (F) Representative images of wound healing in the scratch assay, showcasing the migratory response of human EPC. (G) Characterization of circulating exosomes from rat peripheral blood. (H) Quantitative analysis of exosomes derived from rat peripheral blood revealed no statistically significant difference in the number of exosomes isolated from equal volumes of circulating blood among all groups ( n = 3 for each group). (I) Cell proliferation assays revealed that exosomes derived from the HC group exhibited a diminished capacity to promote EPC proliferation compared to those from the NC group in rats. In contrast, exosomes induced by 8 weeks of aerobic exercise significantly enhanced EPC proliferation ( n : 5–6 for each group). * p < 0.05, HC vs . NC; ## p < 0.01, HE vs . HC. (J) Scratch assays indicated that exosomes derived from the HC group exhibited a diminished capacity to enhance EPC migration rates compared to those from the NC group in rats. In contrast, exosomes induced by 8 weeks of aerobic exercise significantly enhanced EPC migration rates ( n = 4 for each group). ** p < 0.01, HC vs . NC; ## p < 0.01, HE vs . HC. (K) Representative images of wound healing in the scratch assay, showcasing the migratory response of rat EPC. CCK-8 = cell counting kit-8; CD63 = cluster of differentiation 63; EPC = endothelial progenitor cells; HC = the high-fat diet with sedentary group; HE = the high-fat diet with exercise group; NC = the normal diet with sedentary group; TSG101 = tumor susceptibility gene 101.

Article Snippet: The primary antibodies used included PI3K (SC-365290, 1:1000; Santa Cruz Biotechnology, Dallas, TX, USA), Akt1 (SC-5298, 1:1000; Santa Cruz), p-Akt (Ser473) (66444-1-lg, 1:1000; Proteintech Group, Rosemont, IL, USA), phosphatase and tensin homolog (PTEN) (60300-1-Ig, 1:1000; Proteintech), tumor susceptibility gene 101 (TSG101) (DF8427, 1:1000; Affinity Biosciences, Cincinnati, OH, USA), cluster of differentiation 63 (CD63) (AF5117, 1:1000; Affinity), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (GB15002-100, 1:4000; Servicebio).

Techniques: Migration, Transmission Assay, Electron Microscopy, Derivative Assay, Isolation, Control, Proliferation Assay, CCK-8 Assay, Wound Healing Assay, Cell Counting

Journal: Journal of Nanobiotechnology

Article Title: Fisetin-loaded nanoparticles as a novel approach for cholesterol regulation in hypercholesterolemia: targeting the ASGR1-mediated mTORC1/AMPK pathway

doi: 10.1186/s12951-026-04181-z

Figure Lengend Snippet: Fisetin reduced the co-localization between ASGR1 and lysosomes and regulated mTORC1/AMPK-BRCA1/BARD1 pathway. ( A ) Representative immunofluorescence images and statistical results of ASGR1 and CD63 protein expression in AML12 cells. Scale bar (20 μm), magnification (400 ×). ( B-G ) Western blot analysis of p-mTOR, p-S6K, p-AMPK, BRCA1 and BARD1 in AML12 cells. Data are mean ± SD, n = 3. * p < 0.05, ** p < 0.01

Article Snippet: Following permeabilization with 0.1% Triton X-100 for 10 min, the cells were blocked with 5% BSA for 1 h. Next, the cells were incubated with primary antibodies against ASGR1 (Proteintech, Wuhan, China, 11739-1-AP, 1:200) and Cluster of Differentiation 63 (CD63) (Abmart, Shanghai, China, M051014, 1:200) for 2 h. Subsequently, the cells were incubated with FITC-conjugated Goat Anti-Mouse IgG (Abbkine, Wuhan, China, A22110, 1:500) and TRITC-conjugated Goat Anti-Rabbit IgG (Zenbio, Chengdu, China, 511202, 1:500) for 1.5 h. The nuclei were then stained with 1 μg/mL DAPI for 10 min.

Techniques: Immunofluorescence, Expressing, Western Blot

Journal: BMC Psychiatry

Article Title: Normalization strategies for protein quantification in plasma astrocyte-derived extracellular vesicles: a clinical applicability study

doi: 10.1186/s12888-026-07796-6

Figure Lengend Snippet: Distribution of particle counts and EV biomarker levels in plasma ADEVs. ( A ) Distribution of ADEV particle counts measured by nano-flow cytometry (NFCM) and nanoparticle tracking analysis (NTA). ( B ) Distribution of five EV biomarkers (CD9, CD63, CD81, TSG101, and Alix) in ADEVs

Article Snippet: The levels of CD9 (Cat# CSB-EL004969HU, CUSABIO, China), CD63 (Cat# CSB-E14107h-IS, CUSABIO, China), CD81 (Cat# CSB-EL004960HU-IS, CUSABIO, China), Alix (Cat# CSB-EL017673HU, CUSABIO, China), TSG101 (Cat# CSB-EL025125HU, CUSABIO, China), and BDNF (Cat# E-EL-H0010, Elabscience, China) in ADEVs were measured using enzyme-linked immunosorbent assay (ELISA) kits.

Techniques: Biomarker Discovery, Clinical Proteomics, Flow Cytometry

Journal: Applied Sciences

Article Title: Development of Automated Exosome Isolation Method Using Epigallocatechin Gallate-Modified Magnetic Beads: Standardized Protocols for Diverse Biofluids

doi: 10.3390/app15116170

Figure Lengend Snippet: Figure 9. Comparative analysis of CD63 expression in exosomes isolated from various types of biological samples using Western blot and ELISA: (A) urine, (B) plasma, (C) serum, and (D) saliva.

Article Snippet: For urine and saliva samples, a human CD63 ELISA kit (Cusabio, Wuhan, China) was used following the manufacturer’s instructions.

Techniques: Expressing, Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Clinical Proteomics