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anti-cd63 lamp-3 (Developmental Studies Hybridoma Bank)

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Developmental Studies Hybridoma Bank anti-cd63 lamp-3
Anti Cd63 Lamp 3, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 95/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+cd63/anti-CD63+LAMP-3/custom%40h5c6%4010%2E64898%2F2026%2E07%2E12%2E738053
Average 95 stars, based on 48 article reviews

anti-cd63 lamp-3 - by Bioz Stars, 2026-09

95/100 stars

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other:

Article Title: Bulk serum extracellular vesicles from stressed mice show a distinct proteome and induce behavioral and molecular changes in naive mice.
Article Snippet: Protein quantification, Western blots and antibodies Protein content was quantified using the Bradford dye-binding method (Bio-Rad Laboratories).

Article Title: African swine fever virus transmembrane protein pEP84R guides core assembly.
Article Snippet: The sources of the mouse mAbs for host cell markers and tag epitopes were as follows: anti- β-Actin (clone AC-15, Sigma), anti-CD63 (H5C6, DSHB, University of Iowa), anti-PDI (1D3, Enzo Life Sciences), anti-ERGIC-53 (OTI1A8, Enzo Life Sciences); anti-GM130 (35/GM130, BD Transduction Laboratories), anti-DNA (IgM Ac 30–10, Sigma), anti-Flag (M2, Sigma), anti-V5 (clone V5-10, Sigma).

Article Title: Anti-liver fibrotic effects of small extracellular vesicle microRNAs from human umbilical cord-derived mesenchymal stem cells and their differentiated hepatocyte-like cells.
Article Snippet: Objective The aim of this study is to identify therapeutic cargos within mesenchymal stem cell (MSC)-derived small extracellular vesicles (sEVs) for the treatment of liver fibrosis, a condition that poses significant health risks.. Results sEVs from human umbilical cordderived MSCs (UCMSCs) and their differentiated hepatocyte-like cells (hpUCMSCs) were found to alleviate liver fibrosis in mouse models, reduce fibrogenic gene expression in the liver, and inhibit hepatic stellate cell (HSC) activation, a central driver of liver fibrosis, in vitro.. Deep sequencing identified differentially abundant microRNAs (miRNAs) (high-abundance: 57, low-abundance: 22) in both UCMSCand hpUCMSC-derived sEVs, compared to HeLa cell-derived sEVs, which lack anti-liver fibrotic activity.

Labeling:

Article Title: Rational Design of New Biocompatible Near-Infrared Conjugated Polymer Nanoparticles for Biomedical Applications
Article Snippet: .. The primary antibodies used were anti-LAMP1 and anti-CD63 (both from DSHB), and fluorescently labeled secondary antibodies were purchased from Jackson ImmunoResearch. ..

Bioprocessing:

Article Title: Chemo‐small extracellular vesicles released in cisplatin‐resistance ovarian cancer cells are regulated by the lysosomal function
Article Snippet: .. The following monoclonal antibodies were used: anti‐ALIX (cat# sc‐53540), anti‐RAB7 (cat# sc‐376362), anti‐RAB11A (cat# sc‐166912) and anti‐β‐actin (cat# sc‐47778) from Santa Cruz Biotechnology (Dallas, TX, USA); anti‐CD63 (cat# ab8219), anti‐GRP94 (cat# ab90458), anti‐RAB22A (cat# ab137093), anti‐TSG101 (cat# ab83) and anti‐ULK1 (cat#ab128859) from Abcam (Cambridge, UK); anti‐LAMP1 clone H4A3 (cat# DSHB‐H4A3) from Developmental Studies Hybridoma Bank (Iowa City, IA, USA); anti‐GM130 (cat# 610823) and anti‐TSG101 (cat# 612696) from BD Bioscience (Becton, NJ, USA); anti‐Syntenin 1 (cat# NBP2‐76873) from Novous Biological (Littleton, Ontario, Canada); anti‐ULK1 S317 (cat# 127535) from Cell Signaling Technology (Danvers, MA, USA). .. The following polyclonal antibodies: anti‐Cathepsin D (cat# AF1014) from R&D Systems (Minneapolis, MN, USA); anti‐CD63 (cat# AM11837) from Ango (San Ramon, CA, USA); anti‐HGS (HRS) (cat# ab15539) from Abcam; anti‐LAMP2A clone AMC2 (cat# 51–2200) from GibcoTM ThermoFisher Scientific (Waltham, MA, USA); anti‐RAB35 (cat# 9690) from Cell Signaling Technology; anti‐RAB27A (cat# 168013) from Synaptic Systems (Göttingen, Germany).

Western Blot:

Article Title: The Parkinson’s Disease related mutant VPS35 (D620N) amplifies the LRRK2 response to endolysosomal stress
Article Snippet: .. Antibodies and other reagents are as follows: anti-actin (1:10 000 WB, Proteintech; 66009-1-IG), anti-actin (1:2000 WB, Proteintech; 20536-1-AP), anti-actin (1:2000 WB, Sigma–Aldrich; A2066), anti-ALIX (1:1000 WB, 1:500 IF, Santa Cruz Biotechnology; sc-53540), anti-Cathepsin D (1:2000 WB, Calbiochem; 219361), anti-CD63 (1:500 IF, Developmental Studies Hybridoma Bank H5C6), anti-CHMP2B (1:500 IF, abcam ab157208), anti-CIM6PR (1:1000 WB, 1:500 IF, gift from Paul Luzio), anti-EEA1 (1:500 IF, BD Biosciences; 610456), anti-FAM21C (1:1000 IF, Merck; ABT79), anti-HA (1:1000 WB, Covance; MMS-101P), anti-HA (1:400 IF, Novus Biotechne; NB600-362), anti-HA (1:500 IF, Roche; 11867423001), anti-LAMP1 (1:1000 WB, 1:500 IF, Cell Signalling Technology; 9091), anti-LAMP1 (1:200 IF, Developmental Studies Hybridoma Bank H4A3), anti-LC3 (1:200 WB, 1:200 IF, Nanotools; 5F10), anti-p62 (1:1000 WB, BD Biosciences; 610833), anti-phosphoRab10 T73 (1 mg/ml WB, abcam; ab230261), anti-phosphoRab12 S106 (1:1000 WB, abcam ab256487), anti-Rab10 total (1:1000 WB, Cell Signalling Technology; 8127), anti-Rab12 total (1 mg/ml WB, MRC PPU SA227), anti-Sortilin (1:1000 WB, 1:1000 IF, abcam; ab16640), anti-TGN46 (1:500 IF, Bio-Rad; AHP500), anti-VPS26A IF, WB (1:1000 WB, 1:800 IF, abcam; ab23892), anti-VPS35 (1:1000 WB, abcam; ab10099), anti-WASHC1 (1:2000 WB, Sigma–Aldrich HPA002689), apilimod dimesylate (Tocris Bioscience; 7283), blasticidin S (Invitrogen; R21001), chloroquine diphosphate salt (Sigma–Aldrich; C6628), concanamycin A (Sigma–Aldrich; C9705), doxycycline hyclate (Sigma–Aldrich; R25005), G-418 solution (Roche; Figure 5. ..

Article Title: The Parkinson’s Disease related mutant VPS35 (D620N) amplifies the LRRK2 response to endolysosomal stress
Article Snippet: .. Antibodies and other reagents are as follows: anti-actin (1:10,000 WB, Proteintech; 66009-1-IG), anti-actin (1:2,000 WB, Proteintech; 20536-1-AP), anti-actin (1:2,000 WB, Sigma-Aldrich; A2066), anti-ALIX (1:1,000 WB, 1:500 IF, Santa Cruz Biotechnology; sc-53540), anti-Cathepsin D (1:2,000 WB, Calbiochem; 219361), anti-CD63 (1:500 IF, Developmental Studies Hybridoma Bank H5C6), anti-CHMP2B (1:500 IF, abcam ab157208), anti-CIM6PR (1:1,000 WB, 1:500 IF, gift from Paul Luzio), anti-EEA1 (1:500 IF, BD Biosciences; 610456), anti-FAM21C (1:1,000 IF, Merck; ABT79), anti-HA (1:1000 WB, Covance; MMS-101P), anti-HA (1:400 IF, Novus Biotechne; NB600-362), anti-HA (1:500 IF, Roche; 11867423001), anti-LAMP1 (1:1,000 WB, 1:500 IF, Cell Signalling Technology; 9091), anti-LAMP1 (1:200 IF, Developmental Studies Hybridoma Bank H4A3), anti-LC3 (1:200 WB, 1:200 IF, Nanotools; 5F10), anti-p62 (1:1,000 WB, BD Biosciences; 610833), anti-phosphoRab10 T73 (1 μ g/ml WB, abcam; ab230261), anti-phosphoRab12 S106 (1:1,000 WB, abcam ab256487), anti-Rab10 total (1:1000 WB, Cell Signalling Technology; 8127), anti-Rab12 total (1 μ g/ml WB, MRC PPU SA227), anti-Sortilin (1:1,000 WB, 1:1,000 IF, abcam; ab16640), anti-TGN46 (1:500 IF, Biorad; AHP500), anti-VPS26A IF, WB (1:1,000 WB, 1:800 IF, abcam; ab23892), anti-VPS35 (1:1000 WB, abcam; ab10099), anti-WASHC1 (1:2,000 WB, Sigma-Aldrich HPA002689), apilimod dimesylate (Tocris Bioscience; 7283), blasticidin S (Invitrogen; R21001), chloroquine diphosphate salt (Sigma-Aldrich; C6628), concanamycin A (Sigma-Aldrich; C9705), doxycycline hyclate (Sigma-Aldrich; R25005), G-418 solution (Roche; 4727878001), L-Leucyl-L-Leucine methyl ester (LLOMe) hydrochloride (Cayman Chemical; CAY16008), MLi-2 (Tocris Bioscience; 5756), nigericin sodium salt (Sigma-Aldrich; N7143), puromycin dihydrochloride (Sigma-Aldrich; P7255), Zeocin (Invitrogen; R25005). .. hTERT-RPE1-Flp-In-Parental (hTERT-RPE1-FRT-TREX, kind gift from Jon Pines, ICR, London) cells were cultured in Dulbecco’s modified Eagle medium DMEM/F12 with GlutaMAX (Gibco; 31331093) supplemented with 10% FBS (Gibco; 10270106) and 1% non-essential amino acids (Gibco; 11150035). hTERT-RPE1-Flp-In-HA-VPS35 cells were maintained as above in the presence of 200 μ g/ml G418 (Sigma; 4727878001) and 5 μ g/ml blasticidin (Thermo Fisher Scientific; R21001).

Article Title: The Parkinson's disease related mutant VPS35 (D620N) amplifies the LRRK2 response to endolysosomal stress
Article Snippet: .. Antibodies and other reagents are as follows: anti-actin (1:10 000 WB, Proteintech; 66009-1-IG), anti-actin (1:2000 WB, Proteintech; 20536-1-AP), anti-actin (1:2000 WB, Sigma–Aldrich; A2066), anti-ALIX (1:1000 WB, 1:500 IF, Santa Cruz Biotechnology; sc-53540), anti-Cathepsin D (1:2000 WB, Calbiochem; 219361), anti-CD63 (1:500 IF, Developmental Studies Hybridoma Bank H5C6), anti-CHMP2B (1:500 IF, abcam ab157208), anti-CIM6PR (1:1000 WB, 1:500 IF, gift from Paul Luzio), anti-EEA1 (1:500 IF, BD Biosciences; 610456), anti-FAM21C (1:1000 IF, Merck; ABT79), anti-HA (1:1000 WB, Covance; MMS-101P), anti-HA (1:400 IF, Novus Biotechne; NB600-362), anti-HA (1:500 IF, Roche; 11867423001), anti-LAMP1 (1:1000 WB, 1:500 IF, Cell Signalling Technology; 9091), anti-LAMP1 (1:200 IF, Developmental Studies Hybridoma Bank H4A3), anti-LC3 (1:200 WB, 1:200 IF, Nanotools; 5F10), anti-p62 (1:1000 WB, BD Biosciences; 610833), anti-phosphoRab10 T73 (1 μg/ml WB, abcam; ab230261), anti-phosphoRab12 S106 (1:1000 WB, abcam ab256487), anti-Rab10 total (1:1000 WB, Cell Signalling Technology; 8127), anti-Rab12 total (1 μg/ml WB, MRC PPU SA227), anti-Sortilin (1:1000 WB, 1:1000 IF, abcam; ab16640), anti-TGN46 (1:500 IF, Bio-Rad; AHP500), anti-VPS26A IF, WB (1:1000 WB, 1:800 IF, abcam; ab23892), anti-VPS35 (1:1000 WB, abcam; ab10099), anti-WASHC1 (1:2000 WB, Sigma–Aldrich HPA002689), apilimod dimesylate (Tocris Bioscience; 7283), blasticidin S (Invitrogen; R21001), chloroquine diphosphate salt (Sigma–Aldrich; C6628), concanamycin A (Sigma–Aldrich; C9705), doxycycline hyclate (Sigma–Aldrich; R25005), G-418 solution (Roche; 4727878001), LLOMe hydrochloride (Cayman Chemical; CAY16008), MLi-2 (Tocris Bioscience; 5756), nigericin sodium salt (Sigma–Aldrich; N7143), puromycin dihydrochloride (Sigma–Aldrich; P7255), Zeocin (Invitrogen; R25005). .. Antibodies and other reagents are as follows: anti-actin (1:10 000 WB, Proteintech; 66009-1-IG), anti-actin (1:2000 WB, Proteintech; 20536-1-AP), anti-actin (1:2000 WB, Sigma–Aldrich; A2066), anti-ALIX (1:1000 WB, 1:500 IF, Santa Cruz Biotechnology; sc-53540), anti-Cathepsin D (1:2000 WB, Calbiochem; 219361), anti-CD63 (1:500 IF, Developmental Studies Hybridoma Bank H5C6), anti-CHMP2B (1:500 IF, abcam ab157208), anti-CIM6PR (1:1000 WB, 1:500 IF, gift from Paul Luzio), anti-EEA1 (1:500 IF, BD Biosciences; 610456), anti-FAM21C (1:1000 IF, Merck; ABT79), anti-HA (1:1000 WB, Covance; MMS-101P), anti-HA (1:400 IF, Novus Biotechne; NB600-362), anti-HA (1:500 IF, Roche; 11867423001), anti-LAMP1 (1:1000 WB, 1:500 IF, Cell Signalling Technology; 9091), anti-LAMP1 (1:200 IF, Developmental Studies Hybridoma Bank H4A3), anti-LC3 (1:200 WB, 1:200 IF, Nanotools; 5F10), anti-p62 (1:1000 WB, BD Biosciences; 610833), anti-phosphoRab10 T73 (1 μg/ml WB, abcam; ab230261), anti-phosphoRab12 S106 (1:1000 WB, abcam ab256487), anti-Rab10 total (1:1000 WB, Cell Signalling Technology; 8127), anti-Rab12 total (1 μg/ml WB, MRC PPU SA227), anti-Sortilin (1:1000 WB, 1:1000 IF, abcam; ab16640), anti-TGN46 (1:500 IF, Bio-Rad; AHP500), anti-VPS26A IF, WB (1:1000 WB, 1:800 IF, abcam; ab23892), anti-VPS35 (1:1000 WB, abcam; ab10099), anti-WASHC1 (1:2000 WB, Sigma–Aldrich HPA002689), apilimod dimesylate (Tocris Bioscience; 7283), blasticidin S (Invitrogen; R21001), chloroquine diphosphate salt (Sigma–Aldrich; C6628), concanamycin A (Sigma–Aldrich; C9705), doxycycline hyclate (Sigma–Aldrich; R25005), G-418 solution (Roche; 4727878001), LLOMe hydrochloride (Cayman Chemical; CAY16008), MLi-2 (Tocris Bioscience; 5756), nigericin sodium salt (Sigma–Aldrich; N7143), puromycin dihydrochloride (Sigma–Aldrich; P7255), Zeocin (Invitrogen; R25005).

Image Search Results

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.

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

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.

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