exosome surface marker proteins cd9  (Proteintech)

Journal: bioRxiv

Article Title: miR-146a-3p packaged in small extracellular vesicles triggers fetal membrane inflammation in response to viral dsRNA through activation of Toll-like Receptor 7 and 8

doi: 10.1101/2025.07.29.667498

Figure Lengend Snippet: (A) sEVs were isolated from the supernatants of FM explants treated for 24 hour with either NT or Poly(I:C). Image shows sEV expression of the surface marker CD9 and CD81 as determined by Western Blot. Chart shows sEV concentrations as determined by NTA. (B) RNA was isolated from sEVs and measured for the TLR7/8-activating miRs, miR-146a-3p, miR-21a, miR-29a and Let7b by RT-qPCR (n=4-8; * p <0.05; # p =0.05).

Article Snippet: The isolated sEV pellet was either: 1) resuspended in 50μl of phosphate buffered saline (PBS, Thermo Fisher Scientific, Waltham, MA) for subsequent nanoparticle tracking analysis and quantification using a Zetaview (Particle Metrix, Ammersee, Germany) set to room temperature, a laser wavelength of 488nm, a sensitivity of 80, and a shutter speed of 100; 2) directly lysed for RNA extraction using the using the SeraMir Exosome RNA Purification Column Kit (Systems Biosciences) as previously described ; or 3) directly lysed into sample buffer for Western blot analysis of the surface protein marker CD9 (#13174, Cell Signaling Technology; Danvers, MA) and CD81 (#56039T, Cell Signaling Technology).

Techniques: Isolation, Expressing, Marker, Western Blot, Quantitative RT-PCR

Journal: Biological Research

Article Title: Dorsal root ganglion-derived exosomes deteriorate neuropathic pain by activating microglia via the microRNA-16-5p/HECTD1/HSP90 axis

doi: 10.1186/s40659-024-00513-1

Figure Lengend Snippet: DRG-Exo are involved in NP in SNL-induced mice. a Flow chart of mouse SNL modeling and DRG-Exo injection. Assessment of mechanical allodynia response in SNL mice using 0.07 g ( b ) or 0.4 g ( c ) von Frey filaments at different time points before and after SNL surgery, respectively. d The responses of SNL mice to thermal hyperalgesia were evaluated at different time points before and after SNL surgery. e The responses of SNL mice to cold hyperalgesia were assessed at different time points before and after SNL surgery, respectively. f Inflammatory infiltration in the spinal cord (L4) of SNL mice was observed using HE staining. g The morphology of DRG-Exo was observed using TEM. h The diameter of DRG-Exo was analyzed using NTA. i Surface markers CD9, CD63, CD81 expression and contaminant proteins Calnexin and GM130 in DRG-Exo using western blot. Assessment of mechanical allodynia response in PBS- or DRG-Exo-treated mice using 0.07 g ( j ) or 0.4 g ( k ) von Frey filaments at different time points before and after SNL surgery, respectively. l The responses of PBS- or DRG-Exo-treated mice to thermal hyperalgesia were evaluated at different time points before and after SNL surgery. m The responses of PBS- or DRG-Exo-treated mice to cold hyperalgesia were assessed at different time points before and after SNL surgery, respectively. n Inflammatory infiltration in the spinal cord (L4) of PBS- or DRG-Exo-treated mice observed using HE staining (arrows represent immune cells and the circle represents the enrichment of immune cells). Values are expressed as mean ± SD. ** p < 0.01 vs the sham group and # p < 0.05, ## p < 0.01 vs the SNL + PBS group by two-way ANOVA

Article Snippet: Total exosomal protein was measured by the bicinchoninic acid (BCA) protein concentration method, and exosomal surface marker proteins CD9 (1:1000, ab307085, Abcam, Cambridge, UK), CD63 (1:1000, ab216130, Abcam), CD81 (1:1000, ab232390, Abcam), and the contaminants calnexin (1:5000, PA5-34754, Thermo Fisher) and GM130 (1:1000, PA5-95727, Thermo Fisher) in 200 μL solution containing Exo were detected using western blot.

Techniques: Injection, Staining, Expressing, Western Blot

Journal: Biological Research

Article Title: Dorsal root ganglion-derived exosomes deteriorate neuropathic pain by activating microglia via the microRNA-16-5p/HECTD1/HSP90 axis

doi: 10.1186/s40659-024-00513-1

Figure Lengend Snippet: The scheme illustrates the mechanism of DRG-Exo in NP (by Figdraw, https://www.figdraw.com/static/index.html ). Exosomal delivery of miR-16-5p by DRG neurons interacts with HECTD1 to regulate ubiquitination levels of HSP90, thereby promoting microglial activation in NP

Article Snippet: Total exosomal protein was measured by the bicinchoninic acid (BCA) protein concentration method, and exosomal surface marker proteins CD9 (1:1000, ab307085, Abcam, Cambridge, UK), CD63 (1:1000, ab216130, Abcam), CD81 (1:1000, ab232390, Abcam), and the contaminants calnexin (1:5000, PA5-34754, Thermo Fisher) and GM130 (1:1000, PA5-95727, Thermo Fisher) in 200 μL solution containing Exo were detected using western blot.

Techniques: Activation Assay