Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Exosomes derived from syncytia induced by SARS-2-S promote the proliferation and metastasis of hepatocellular carcinoma cells

doi: 10.3389/fcimb.2024.1415356

Figure Lengend Snippet: Analysis of exosome purity. (A) Illustration of the isolation of Ctr-Exos and Syn-Exos. (Created with BioRender.com). (B) Phenotypic analysis of Ctr-Exos and Syn-Exos using NTA. The X-axis represents the vesicle diameter, and the Y-axis represents the vesicle concentration (particles/ml). (C) WB analysis was performed to examine typical exosomal biomarkers (CD9, CD63 and TSG101) in Ctr-Exos and Syn-Exos. (D) Representative SEM images of Ctr-Exos and Syn-Exos.

Article Snippet: Anti-CD63 (bsm-52384R, 1:500 dilution) was purchased from Bioss (Beijing, China).

Techniques: Isolation, Concentration Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Higher Urine Exosomal miR-193a Is Associated With a Higher Probability of Primary Focal Segmental Glomerulosclerosis and an Increased Risk of Poor Prognosis Among Children With Nephrotic Syndrome

doi: 10.3389/fcell.2021.727370

Figure Lengend Snippet: Representative micrographs show renal CD63 staining (IHC, 400×). The expression level of CD63 was significantly up-regulated in patients with FSGS compared with TBMD, which was mainly located in the cytoplasm of podocytes within non-sclerotic tuft segments (indicated by black arrows).

Article Snippet: For immunohistochemical staining, we incubated the sections with rabbit anti-human CD63 antibodies (1:200, Wuhan Boster Biological Technology Co., Ltd, China), and then dewaxed, rehydrated, retrieved antigen, inactivated sections with 3% H 2 O 2 , and blocked non-specific binding with 10% goat sera.

Techniques: Staining, Expressing

Journal: Cells

Article Title: DICAM in the Extracellular Vesicles from Astrocytes Attenuates Microglia Activation and Neuroinflammation.

doi: 10.3390/cells11192977

Figure Lengend Snippet: Figure 4. DICAM is highly enriched in the exosome from stimulated primary astrocytes. (A) Serum DICAM levels from patients with CRPS and age- and sex-matched healthy controls were determined using ELISA analysis and compared by Mann–Whitney U test. n = 5 per group. (B) RT-PCR analysis for DICAM, GFAP (a marker for astrocyte), and CD11b (a marker for microglia) were performed with mRNA isolated from primary astrocytes, primary microglia, and murine microglia cell-line BV-2. (C) The protein level of DICAM in the culture supernatants of WT primary astrocytes was measured using ELISA analysis. Primary astrocytes were treated with 100 ng/mL of LPS and 10 ng/mL of IFN-γ for 24 h. * p < 0.05 by Mann–Whitney U test. n = 3 per group. (D) Representative Western blot image of the total cell lysates and exosomes secreted by primary astrocytes in the medium showing DICAM and CD63 expressions (a marker for exosome).

Article Snippet: Primary antibodies against p38 MAPK, p-p38 MAPK, JNK, p-JNK, ERK1/2, p-ERK1/2, Akt, p-Akt, IκBα, p-IκBα, p65, p-p65, Stat3, and p-Stat3 were purchased from Cell Signaling Technology (Beverly, MA, USA); antibodies against CD14, CD68, and CD63 were purchased from Cusabio Technology (Houston, TA, USA); the ERK antibody was purchased from BD Biosciences (Franklin lakes, NJ, USA); and the β-actin antibody was purchased from Sigma Aldrich (St. Louis, MO, USA).

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Reverse Transcription Polymerase Chain Reaction, Marker, Isolation, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: microRNA Expression of Renal Proximal Tubular Epithelial Cells and Their Extracellular Vesicles in an Inflammatory Microenvironment In Vitro

doi: 10.3390/ijms241311069

Figure Lengend Snippet: Characterization of PTCs ( A , B ) and EVs isolated from PTC supernatant ( C , D ). ( A ) Characteristic phase contrast microscopy of confluent PTCs cultured in standard cell culture. ( B ) Representative flow cytometric overlay histograms of characteristic PTC marker expression (CD13, CD26, and EpCAM), and of CD63, a characteristic marker of exosomes. The black histograms represent isotype controls. ( C ) Representative nanoparticle tracking analysis (NTA) of EVs isolated from unstimulated PTC after standard cell culture for 48 h. ( D ) Representative flow cytometric overlay histogram of PTC-EVs isolated using EpCAM-beads and immunostained with CD63-APC. The black histogram represents an unstained control. ( E ) Representative nanoparticle tracking analysis (NTA) of EVs isolated from PTCs after culture in an inflammatory microenvironment for 48 h.

Article Snippet: The bead-EV solution (100 µL) was then stained with 20 µL of APC-labeled anti-CD63 detection antibody (ImmunoTools, Friesoythe, Germany) and incubated under rotation at RT in the dark for 45 min and washed and re-suspended in a 300 µL isolation buffer.

Techniques: Isolation, Microscopy, Cell Culture, Marker, Expressing