Journal: International journal of molecular sciences

Article Title: Thermostable Basic Fibroblast Growth Factor Enhances the Production and Activity of Human Wharton's Jelly Mesenchymal Stem Cell-Derived Extracellular Vesicles.

doi: 10.3390/ijms242216460

Figure Lengend Snippet: Figure 3. Characterization of WJ-MSC-derived exosomes treated with WT-bFGF (WT-3D EVs) or TS-bFGF (TS-3D EVs). (A) The particle size comparison was carried out using ZetaView. The concen- tration is indicated as particles/mL. Two-dimensional EV and 3D EV: EVs derived from WJ-MSCs cultured under 2D or 3D conditions, respectively. (B) Comparison of exosome production. Increased exosome production was observed with TS-bFGF treatment compared to WT-bFGF treatment under the same culture conditions. (C) Expression analysis of CD63 and CD9 (positive exosome mark- ers) and GM130 and calnexin (negative exosome markers) using Western blotting. CD63 and CD9 expressions were detected in all exosome groups. (D) Analysis of CD81 expression in exosomes isolated using exosome human CD9 beads. *** p < 0.01, **** p < 0.001 compared to the control group. #### p < 0.0001 compared to the WT-3D EV group.

Article Snippet: After washing thrice with filtered PBS, the secondary antibodies were CD63-PE (556020, BD Pharmingen) and CD81-APC (130–119-787, Miltenyi Biotec).

Techniques: Derivative Assay, Comparison, Cell Culture, Expressing, Western Blot, Isolation, Control

Journal: Bioactive materials

Article Title: Tumor-derived apoptotic extracellular vesicle-mediated intercellular communication promotes metastasis and stemness of lung adenocarcinoma.

doi: 10.1016/j.bioactmat.2024.02.026

Figure Lengend Snippet: Fig. 1. Characteristics of apoptotic extracellular vesicles (apoEVs) derived from LUAD cells and normal lung epithelial cells. (A) Schematic diagram of apoEVs isolation. A549 and BEAS2b are induced by STS or Cisplatin (DDP) for 24 h and then apoptotic cell suspensions are isolated using a differential centrifugation to obtain apoEVs. (B) Representative flow cytometry analysis of BEAS2b apoptosis after STS treatment, and A549 apoptosis after Cisplatin (DDP) and STS treatment. (C) Representative TEM micrographs of apoEVs collected from the conditioned medium after apoptosis induction treatment of A549 and BEAS2b cells evaluated are shown. Scale bar, 200 nm. (D) Size distribution of BEAS2b- and A549-apoEVs were measured by nanoparticle tracking analyzer (NTA). (E) Representative histograms show the profile of the CD9, CD63 and CD81 levels in comparison to isotype control-stained for the BEAS2b and A549-derived apoEVs. (F) Nanoflow cytometry show apoEVs express apoptotic vesicles-specific surface markers Ptdser (shown by Annexin V staining). (G) Western blot of the indicated proteins in apoEVs, Tubulin (loading control) were used, western blotting was repeated at least three times and representative data are shown.

Article Snippet: ApoEVs derived from LUAD cell lines and Human Normal Lung Epithelial Cells BEAS2b were incubated with PE-conjugated anti-CD9, CD63 and CD81 (Elabscience), the general EV markers, at 4 ◦C for 30 min. To detect phosphatidylserine (Ptdser), FITC-Annexin V was used to stain apoEVs suspended in Annexin V Binding Buffer (Elabscience) for 15 min at 4 ◦C.

Techniques: Derivative Assay, Isolation, Centrifugation, Flow Cytometry, Comparison, Control, Staining, Cytometry, Western Blot

Journal: Scientific Reports

Article Title: Cytosolic nucleic acid sensors of the innate immune system promote liver regeneration after partial hepatectomy

doi: 10.1038/s41598-018-29924-3

Figure Lengend Snippet: Release of CD81-positive extracellular vesicles during liver regeneration. Exosomes were isolated from serum samples using ExoQuick and exosome lysates were analysed by Western blotting using an antibody against CD81 (A, upper panel). The protein band corresponding to CD81 is indicated by an arrowhead. Protein loading was controlled by coomassie blue staining of gels (A, lower panel). For each time point, samples from six individual wildtype mice are shown. Full length gels are depicted. Expression of CD81 was quantified by densitometric analyses ( B ). * P < 0.05, ** P < 0.01 (two-tailed unpaired Student’s t-test or the Mann-Whitney U test).

Article Snippet: Membranes were incubated with the following primary antibodies: STAT3, p-STAT3 (Tyr705), p-RB (S807/811), p-CDK2 (T160), cyclin E1, and GAPDH (all from Cell Signaling Technology, Danvers, MA); β-tubulin and CDK1 (both from Abcam, Cambridge, MA); CD81 (St. John’s Laboratory, London, UK); and cyclin D1 (Santa Cruz Biotechnology, Dallas, TX).

Techniques: Isolation, Western Blot, Staining, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Gemcitabine-cisplatin chemotherapy plus anti-PD-L1 therapy reinvigorates antitumor immune response by reprogramming the intrahepatic cholangiocarcinoma microenvironment

doi: 10.3389/fimmu.2025.1666393

Figure Lengend Snippet: Differential analysis of CD8 + T cells between untreated and GCP-treated ICC samples. (A) UMAP plots showing all T cells labeled in different colors according to cell type annotation. (B) Volcano plot showing the differentially expressed genes of intra-tumoral CD8 + T cells between pretreated and posttreated ICC samples. (C) Immunofluorescence images showing the infiltration of CD8 + CD81 + T cells in treatment-naïve ICC samples from the TMA cohort (n=89). Kaplan–Meier survival curves for OS (D) and RFS (E) of 89 ICC patients grouped by infiltration levels of CD8 + CD81 + T cells. P values were determined via log-rank test. (F) Boxplot showing infiltration levels of CD8 + CD81 + T cells in GCP-treated tumors revealed by multiplex immunofluorescence in GCP cohort. (G) Violin plots showing the expression scores of tissue resident, exhausted, and co-stimulatory gene signatures in tumor-infiltrating CD8 + T cells between pretreated and posttreated samples. The P values were calculated by Wilcox test. OS, overall survival; RFS, recurrence-free survival. ****p < 0.0001.

Article Snippet: After that, the slides were incubated with CD81 primary antibody (1:200 dilution, A01281-2, BOSTER) and Alexa Fluor 488 donkey anti-rabbit antibody as described above.

Techniques: Labeling, Immunofluorescence, Multiplex Assay, Expressing