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: Signal Transduction and Targeted Therapy

Article Title: Breast cancer-associated fibroblast-derived exosomal miR-1290 promotes triple-negative breast cancer cell proliferation

doi: 10.1038/s41392-026-02922-y

Figure Lengend Snippet: miR-1290 is upregulated in breast cancer and BCAF/MB-231-derived exosomes. a Nanoparticle tracking analysis of exosomes isolated from BCAFs and MB-231 cells showing their size distribution. The mean size and standard deviation (SD) are indicated for each sample. b TEM images of exosomes isolated from BCAFs (upper) and MB-231 cells (lower) showing the characteristic morphology of the exosomes. Scale bar, 100 nm. c Western blot analysis of exosome markers (CD63, CD81, and flotillin-1), CAF markers (α-SMA and FAP) and cellular markers (GM130 and H3) in BCAF cells, MB-231 cells and their derived exosomes. d BCAF-derived exosome uptake by TNBC cells. PKH67-labeled BCAF-derived exosomes (green) were incubated with BT-549 and Hs578T cells for 1 hour. The cell nuclei were stained with Hoechst 33342 (blue). Scale bar, 20 μm. e Heatmap showing miRNA expression profiles in exosomes derived from BCAFs and MB-231 cells. Red indicates high expression, and blue indicates low expression. miR-1290 (highlighted in red) is highly expressed in both exosome types. f Volcano plots showing differentially expressed miRNAs between breast cancer or TNBC samples and normal controls in five independent GEO datasets ( GSE37963 , GSE45666 , GSE45498 , GSE38167 , and GSE61438 ). Red and blue dots represent upregulated and downregulated miRNAs, respectively, selected using fold-change and nominal p value cutoffs (|log2(fold change)| > 0.58 and nominal p < 0.05, two-tailed Welch’s t test). g Venn diagram analysis of upregulated miRNAs across the five GEO datasets revealed four commonly upregulated miRNAs. h Violin plot showing miR-1290 expression levels in serum samples from normal individuals ( N , n = 2686) and breast cancer patients ( P , n = 1280) from the GSE73002 dataset. The expression of miR-1290 was significantly upregulated in the breast cancer patient samples (*** p < 0.001, Mann‒Whitney U test). i Analysis of serum exosomal miR-1290 levels in normal ( N , n = 20) and TNBC patient ( P , n = 39) samples collected at Keimyung University Dongsan Hospital. The results support the potential of miR-1290 as a candidate diagnostic marker in an independent cohort (*** p < 0.001, Mann‒Whitney U test). j Kaplan‒Meier plot of overall survival for TNBC patients based on serum exosomal miR-1290 expression levels from Keimyung University Dongsan Hospital ( p = 0.006, log-rank test)

Article Snippet: Membranes were incubated with the following primary antibodies: CD63 (1:200; ABclonal, A5271), CD81 (1:200; ABclonal, A5270), flotillin-1 (1:1000; ABclonal, A6220), α-SMA (1:1000; ABclonal, A17910), FAP (1:1000; ABclonal, A23789 ), GM130 (1:1000; Abcam, ab52649), H3 (1:1000; Cell Signaling Technology, 4620S), ACTB (1:20,000; ABclonal, AC026), TBC1D4 (1:200; Cell Signaling Technology, 2670S), P70S6K (1:500; Cell Signaling Technology, 9202S), phospho-P70S6K (1:500; ABclonal, AP0564), and Ki67 (1:1000; Abcam, ab16667).

Techniques: Derivative Assay, Exosomes, Analysis, Isolation, Standard Deviation, Morphology, Western Blot, Labeling, Incubation, Staining, Expressing, Two Tailed Test, Blood/Plasma/Serum Samples, Diagnostic Assay, Marker

Journal: Signal Transduction and Targeted Therapy

Article Title: Breast cancer-associated fibroblast-derived exosomal miR-1290 promotes triple-negative breast cancer cell proliferation

doi: 10.1038/s41392-026-02922-y

Figure Lengend Snippet: miR-1290 promotes breast cancer cell proliferation. a Crystal violet staining of BT-549 and Hs578T cells treated with DPBS or BCAF-derived exosomes (left). Quantification of cell numbers (middle). qRT‒PCR analysis of miR-1290 expression in recipient cells (right). b Crystal violet staining of BT-549 and Hs578T cells treated with DPBS- or MB-231-derived exosomes (left). Quantification of cell numbers (middle). qRT‒PCR analysis of miR-1290 expression in recipient cells (right). Scale bar, 500 μm. c Crystal violet staining of BT-549 and Hs578T cells transfected with the miR-1290 mimic or mimic-NC (left). Quantification of cell numbers (middle). qRT‒PCR analysis of miR-1290 expression (right). Scale bar, 500 μm. d Representative images of xenograft tumors derived from BT-549 cells transfected with the miR-1290 mimic or mimic-NC. e Growth curves of xenograft tumors in NOD/Shi-scid/IL-2Rγ null mice inoculated with BT-549 cells transfected with the miR-1290 mimic or mimic-NC ( n = 6 per group). The tumor volume was compared between the two groups at the experimental endpoint. f Tumor weight measurements at the endpoint of the xenograft experiment shown in ( e ) ( n = 6 per group). g Representative immunocytochemistry images showing the localization and expression of Ki67 in BT-549 and Hs578T cells treated with BCAF exosomes or MB-231 exosomes and transfected with the miR-1290 mimic or mimic-NC. The nuclei were counterstained with DAPI. Scale bar, 150 μm. The data are shown as the means ± SDs of three independent experiments unless otherwise indicated, and statistical significance was determined using Student’s t tests (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: Membranes were incubated with the following primary antibodies: CD63 (1:200; ABclonal, A5271), CD81 (1:200; ABclonal, A5270), flotillin-1 (1:1000; ABclonal, A6220), α-SMA (1:1000; ABclonal, A17910), FAP (1:1000; ABclonal, A23789 ), GM130 (1:1000; Abcam, ab52649), H3 (1:1000; Cell Signaling Technology, 4620S), ACTB (1:20,000; ABclonal, AC026), TBC1D4 (1:200; Cell Signaling Technology, 2670S), P70S6K (1:500; Cell Signaling Technology, 9202S), phospho-P70S6K (1:500; ABclonal, AP0564), and Ki67 (1:1000; Abcam, ab16667).

Techniques: Staining, Derivative Assay, Exosomes, Analysis, Expressing, Transfection, Xenograft Assay, Immunocytochemistry

Journal: Signal Transduction and Targeted Therapy

Article Title: Breast cancer-associated fibroblast-derived exosomal miR-1290 promotes triple-negative breast cancer cell proliferation

doi: 10.1038/s41392-026-02922-y

Figure Lengend Snippet: miR-1290 promotes breast cancer cell proliferation by directly targeting TBC1D4. a Venn diagram analysis of genes downregulated according to the RNA-seq data of MB-231 exosome-treated (FPKM ratio < 0.7 versus DPBS; n = 981) and miR-1290 mimic-transfected BT-549 cells (FPKM ratio < 0.5 versus mimic-NC; n = 676) with predicted miR-1290 targets from TargetScan ( n = 4601), identifying 12 common candidate genes (left). Expression of TBC1D4 under the indicated conditions (right). b Expression levels of TBC1D4 in normal ( n = 114) and breast cancer ( n = 1097) tissues from the TCGA database (*** p < 0.001, Mann‒Whitney U test). c Kaplan‒Meier plot of the overall survival of breast cancer patients from the TCGA database. The survival rate of the high-TBC1D4 subgroup was significantly greater than that of the low-TBC1D4 subgroup ( p = 0.003, log-rank test). d qRT‒PCR analysis of TBC1D4 expression in BT-549 and Hs578T cells treated with the indicated conditions. e Western blot analysis of TBC1D4 expression in BT-549 and Hs578T cells under the indicated conditions. ACTB was used as a loading control. f Representative immunocytochemistry images showing the localization and expression of TBC1D4 in BT-549 and Hs578T cells treated with BCAF exosomes and transfected with the miR-1290 mimic or mimic-NC. The nuclei were counterstained with DAPI. Scale bar, 150 μm. g Dual-luciferase reporter assay in HEK293 cells cotransfected with wild-type or mutant TBC1D4 3’-UTR luciferase reporter vectors and miR-1290 mimic or mimic-NC (six technical replicates) (n.s. not significant). h qRT‒PCR analysis of TBC1D4 expression in BT-549 and Hs578T cells transfected with siTBC1D4 or siCont. i Western blot analysis of TBC1D4 expression in BT-549 and Hs578T cells transfected with siTBC1D4 or siCont. ACTB was used as a loading control. j Crystal violet staining of BT-549 and Hs578T cells transfected with siTBC1D4 or siCont and quantification of cell numbers. Scale bar, 500 μm. k Representative immunocytochemistry images showing the localization and expression of Ki67 in BT-549 and Hs578T cells transfected with siTBC1D4 or siCont. The nuclei were counterstained with DAPI. Scale bar, 150 μm. The data are shown as the means ± SDs of three independent experiments, and statistical significance was determined using Student’s t tests (*** p < 0.001)

Article Snippet: Membranes were incubated with the following primary antibodies: CD63 (1:200; ABclonal, A5271), CD81 (1:200; ABclonal, A5270), flotillin-1 (1:1000; ABclonal, A6220), α-SMA (1:1000; ABclonal, A17910), FAP (1:1000; ABclonal, A23789 ), GM130 (1:1000; Abcam, ab52649), H3 (1:1000; Cell Signaling Technology, 4620S), ACTB (1:20,000; ABclonal, AC026), TBC1D4 (1:200; Cell Signaling Technology, 2670S), P70S6K (1:500; Cell Signaling Technology, 9202S), phospho-P70S6K (1:500; ABclonal, AP0564), and Ki67 (1:1000; Abcam, ab16667).

Techniques: Analysis, RNA Sequencing, Transfection, Expressing, Western Blot, Control, Immunocytochemistry, Exosomes, Luciferase, Mutagenesis, Staining

Journal: International Journal of Molecular Sciences

Article Title: Proteasome-Enriched hPPSCs-Derived EVs Attenuate Hypoxic Injury in Endothelial Cells via Proteasome-Mediated HIF-1α Degradation

doi: 10.3390/ijms27157013

Figure Lengend Snippet: Characterization and proteomics of human placenta-derived perivascular stem cells (hPPSCs)–extracellular vesicles (EVs). ( A ) Transmission electron microscopy (TEM) characterization of EVs. Scale bar = 200 nm. The arrow is pointing at the EV. ( B ) Nanoparticle tracking analysis (NTA) of EVs. ( C ) WB analysis of ALIX, CD63, and CD81 in EVs. Equal total proteins were loaded per lane for EV samples from two stem cell types. ( D ) Quantitative analysis of ALIX, CD63, and CD81 protein levels. ( E ) WB analysis of Calnexin in EVs and hPPSCs. ( F ) Volcano plot analysis of differentially expressed proteins (DEPs) between hUCMSCs-EVs and hPPSCs-EVs. Green, red, and blue dots represent downregulated proteins, upregulated proteins, and proteins with no significant changes, respectively. The critical criteria were set to FC > 1.2 or FC < 0.83 (corresponding to |log 2 FC| > 0.263), with p < 0.05. ( G ) Protein domain analysis. ( H , I ) GO functional annotation biological process (BP) and GO functional annotation cell composition (CC) enrichment bubble analysis. The closer to red, the smaller the p -value. ( J ) KEGG enrichment analysis. The abscissa denotes the ratio, while the ordinate stands for the category. n = 3, p < 0.05.

Article Snippet: The antibodies employed for Western blot in this study are listed below: Anti-ALIX antibody (Abcam, ab275377, rabbit, Cambridge, MA, USA), diluted at 1:1000; Anti-CD81 antibody (ABclonal, A5270, rabbit, Woburn, MA, USA), diluted at 1:1000; Anti-CD63 antibody (ABclonal, A19023, rabbit), diluted at 1:1000; Anti-HIF-1α antibody (ABclonal, A16873, rabbit), diluted at 1:1000; Anti-Ubiquitin (P4D1) antibody (SANTA, sc-8017, mouse), diluted at 1:1000; Anti-PSMD11 antibody (ABclonal, A15306, rabbit), diluted at 1:500; Anti-PSMB5 antibody (ABclonal, A1975, rabbit), diluted at 1:1000; Anti-Proteasome 20S alpha + beta antibody (Abcam, ab22673, rabbit), diluted at 1:1000; Anti-GAPDH antibody (ABclonal, A19056, rabbit), diluted at 1:5000; Anti-Calnexin antibody (proteintech, 10427-2-AP, rabbit), diluted at 1:10,000; HRP-linked anti-rabbit IgG (CST, 7074, goat), diluted at 1:10,000; and HRP-linked anti-mouse IgG (CST, 7076, horse), diluted at 1:10,000.

Techniques: Derivative Assay, Transmission Assay, Electron Microscopy, Functional Assay

Journal: Medicine

Article Title: Human amniotic epithelial cell-derived exosomes promote conjunctival goblet cells proliferation and mucin secretion

doi: 10.1097/MD.0000000000049647

Figure Lengend Snippet: Identification and characterization of hAECs-Exo. (A) Morphology of exosomes in the hAECs-Exo group was observed under transmission electron microscopy (Scalar bar = 200 nm). (B) Diameter of exosomes in the hAECs-Exo group was analysis under NTA. (C) Western blot was used to observe the expression of CD63, CD81, and Grp94 proteins in hAECs-Exo. hAECs-Exo = human amniotic epithelial cell-derived exosomes, NTA = nanoparticle tracking analysis.

Article Snippet: The polyvinylidene fluoride membrane was treated with protein-free rapid blocking solution (PS108P, EpiZyme) for 15 minutes, and then incubated with primary antibodies against exosome markers CD63 (1:1000, ABclonal, A19023) and CD81 (1:1000, ABclonal, A4863), as well as the negative marker Grp94 (1:1000, ABclonal, A0989) overnight.

Techniques: Transmission Assay, Electron Microscopy, Western Blot, Expressing, Derivative Assay

Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: Senescent Microenvironment-Educated Mesenchymal Stem Cells Release High-Affinity Senescent NPC Domesticated Extracellular Vesicles. (A) Schematic diagram of the experimental setup for educating MSCs with SASP-CM to generate D-EVs versus N-EVs. (B) Confocal microscopy images showing different EVs internalization by senescent NPCs after 12 h in vitro. (C) Flow cytometry and quantification analysis of different EVs uptake by senescent NPCs. (D) In vivo validation of the senescent niche. Representative fluorescence images following injection of senescence-tracer (Red). (E) In vivo PKH26-labeled D-EVs tracking. (F) Representative SA-β-Gal images and quantification of MSCs treated with SASP-CM or not. (G) Gene Ontology (GO) analysis confirming enrichment of external encapsulating structure organization and cytokine production in Biological Process (BP) categories. (H) Heatmap indicating gene expression associated with EVs biogenesis within D-MSCs and N-MSCs. (I) Heatmap indicating gene expression associated with cytokine production within D-MSCs and N-MSCs. (J and L) Gene Ontology (GO) analysis confirming enrichment of terms related to vesicle organization and transport in the Cellular Component (CC) categories. (K) Western blot analysis confirmed core senescence markers p16 and p21 and DNA damage marker γ-H2AX in N-MSC and D-MSC. (M) Western blot analysis confirmed the expression of CD9, CD63, TSG101, Calnexin, and GM130 in MSC-EVs, N-EVs, or D-EVs. (N) TEM images showing the morphology and size of MSC-derived EVs, N-EVs, and D-EVs. (O) NTA shows size distribution in MSC-EVs, N-EVs, or D-EVs. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibodies against GAPDH (1:5000, 104941-AP, Proteintech), TSG101 (1:1000, DF8427, Affinity), CD9 (1:1000, AF5139, Affinity), CD63 (1:2000, 25682-1-AP, Proteintech), Calnexin (1:5000, 10427-2-AP, Proteintech), GM130 (1:20000, 11308-1-AP, Proteintech), CXCR3 (1:5000, 26756-1-AP, Proteintech), CXCL10 (1:2000, 10937-1-AP, Proteintech), MMP3 (1:2000, 17873-1-AP, Proteintech), ADAMTS5 (DF13268, Affinity), P16 (AF5484, Affinity), P21 (10355-1-AP, Proteintech), GPX4 (1:1000, 381958, Zen-bio), SLC7A11 (1:1000, 26864-1-AP, Proteintech), ACSL4 (1:5000, 22401-1-AP, Proteintech) and Tubulin (1:10000, T40103 , Abmart) overnight at 4 °C.

Techniques: Confocal Microscopy, In Vitro, Flow Cytometry, In Vivo, Biomarker Discovery, Fluorescence, Injection, Labeling, Gene Expression, Western Blot, Marker, Expressing, Derivative Assay