primary antibodies: cd63, cd81, tsg101, calnexin, pi3k, p-pi3k, akt, p-akt, β-actin  (Abmart Inc)

Journal: The FASEB Journal

Article Title: Bone Marrow Mesenchymal Stem Cell‐Derived Exosomes Promote M2 Polarization and Protect Against Acute Lung Injury

doi: 10.1096/fj.202502772R

Figure Lengend Snippet: Preparation of mouse BMSCs and their exosomes. (A) Morphological observation of mouse BMSCs at 120× magnification. To ensure objectivity, five random fields were selected for observation in each analysis. (B) Flow cytometry analysis of the expression of surface markers CD105, CD90, CD73 and CD34 in passage 2 BMSCs. (C) SEM characterization of BMSCs‐Exos. (D) Particle size analysis of BMSCs‐Exos using NTA. (E) Western blot analysis of CD9, CD63, and CD81 protein expression in BMSCs‐Exos, with lysis buffer used as a negative control. Three biological replicates were performed for the experiment ( n = 3), and each replicate was tested three times.

Article Snippet: Rabbit anti‐mouse CD9, CD63, and CD81 primary antibodies (ab223052/ab216130, Abcam; 10 037, CST, MA, USA) were diluted at 1:400, 1:400, and 1:600, respectively.

Techniques: Flow Cytometry, Expressing, Particle Size Analysis, Western Blot, Lysis, Negative Control

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

Journal: International Journal of Molecular Sciences

Article Title: Umbilical Cord Mesenchymal Stem Cell-Derived Extracellular Vesicles Enhance Chondrocyte Function by Reducing Oxidative Stress in Chondrocytes

doi: 10.3390/ijms26167683

Figure Lengend Snippet: Characterization of UCMSCs and UCMSC-EVs. ( A ) Phase-contrast image of UCMSCs cultured on the plastic culture plate. Scale bar: 100 μm. ( B ) Representative histograms showing the expression of CD105, CD90, CD73, CD45, CD34, and HLA-DR on the surface of UCMSCs. Unstained cells were included as negative controls to validate the specificity of antibody staining. ( C ) Flow cytometry analysis to quantify the percentage of UCMSC-EVs positive for CD9, CD63, and CD81. ( D ) Western blot analysis of the protein expressions of CD63, CD81, and α-tubulin in UCMSCs and UCMSC-EVs. ( E ) Morphology of UCMSC-EVs (arrowhead), as observed using TEM.

Article Snippet: Overnight incubation at 4 °C was conducted using primary antibodies against CD81 (Proteintech, Rosemont, IL, USA, 66866-1-lg; 1:1000), CD63, and α-tubulin.

Techniques: Cell Culture, Expressing, Staining, Flow Cytometry, Western Blot