Journal: Frontiers in Cell and Developmental Biology
Article Title: Extracellular vesicles promote activation of pro-inflammatory cancer-associated fibroblasts in oral cancer
doi: 10.3389/fcell.2023.1240159
Figure Lengend Snippet: (A) Identification, quantification and characterization of representative EV particles from Cal 27, SCC-9 and SCC-25 cells according to NanoSight. Size distribution in nm, error bars indicate ± 1 SEM. (B) Protein from OSCC cell and EV protein tested for the vesicle markers Alix, HSP70, Flotillin-1, and CD81 together with GRP94 (negative marker for small EVs) with Western blot. (C) – (G) Flow cytometry analysis of percentage of fibroblasts positive for CAF markers upon co-culture with Cal 27, SCC-9, or SCC-25 derived EVs compared to PBS or TGFβ. (C) FAP, (D) PDPN, (E) CD29, (F) PDGFRβ, and (G) αSMA. n = 4. Statistics: Ordinary one-way ANOVA with Tukey’s multiple comparison test.
Article Snippet: Cells were stained with the following monoclonal antibodies: anti-alpha-Smooth Muscle Actin (αSMA) (Novus Biologicals Cat #NBP2-34760APC), anti- Human Fibroblast Activation Protein alpha (FAP) (R&D Systems Cat #FAB3715P-100), anti-Podoplanin (PDPN) (BioLegend Cat #337006), anti-CD29 (BioLegend Cat #303020), and anti-PDGFRβ (Cell Signaling Technology Cat #3169S) along with Brilliant Violet 421TM Donkey anti-rabbit IgG (BioLegend Cat #406410).
Techniques: Marker, Western Blot, Flow Cytometry, Co-Culture Assay, Derivative Assay, Comparison