cd9  (Danaher Inc)

Journal: Journal of Nanobiotechnology

Article Title: Vascular wall microenvironment: exosomes secreted by adventitial fibroblasts induced vascular calcification

doi: 10.1186/s12951-023-02000-3

Figure Lengend Snippet: Identification and uptake assay of AFs-derived exosomes. A Representative transmission electron microscopic photographs of AFs NPi -Exos and AFs HPi -Exos. The scale bar represents 200 μm. B The diameter distribution of AFs NPi -Exos and AFs HPi -Exos was detected by DLS analysis. C Western blot analysis of exosome-specific markers TSG101, CD9, and CD81 in exosomes secreted by NPi/HPi-induced AFs (AFs NPi -Exos, AFs HPi -Exos). D Representative images of AFs NPi -Exos and AFs HPi -Exos uptake by VSMCs after 12 h incubation. Nuclei stained by DAPI in blue and AFs-Exos stained by PKH26 in red. Scale bar: 50 μm

Article Snippet: After blocking with 5% non-fat milk for 1 h, the membrane was incubated overnight at 4 °C with primary antibody, including CD9 (ab92726, 1:1000, abcam), CD81 (ab109201, 1:1000, abcam), TSG101 (bs-1365R, 1:1000, bioss), Runx2 (ab23981, 1:2000, abcam), BMP2 (bs-10696R, 1:1000, bioss), GAPDH (10494-1-AP, 1:4000, proteintech), BMP4 (bs-1374R, 1:1000, bioss), Crim1 (bs-21654R, 1:1000, bioss), followed by incubation with the horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature.

Techniques: Derivative Assay, Transmission Assay, Western Blot, Incubation, Staining

Journal: Scientific reports

Article Title: Engineered nanovesicles from stromal vascular fraction promote angiogenesis and adipogenesis inside decellularized adipose tissue through encapsulating growth factors.

doi: 10.1038/s41598-022-27176-w

Figure Lengend Snippet: Figure 1. Characteristics of SVF-EVs. (A) TEM images of SVF-EVs. (B) NTA analysis of SVF-EVs. The confirmed size distribution with a range of diameter around 100–300 nm. (C) Western blot shows that SVF- EVs express tetraspanin proteins like TSG101 and CD81. (D) Western blot reveals that SVF-EVs encapsulate proteins such as PPARγ, C/EBPα, Ang-1, and VEGF. (E) Confocal images verify the internalization of PKH26- labeled SVF-EVs (red) into endothelial cell (CD31-labeled, green), and the amount of endocytosed SVF-EVs after co-culturing for 4 h is more than that for 2 h (F).

Article Snippet: Then membranes were blocked with 5% skimmed milk, and incubated with specific antibodies against TSG101 (ab133586, Abcam, UK), CD81(ab109201, Abcam, UK), PPARγ (sc-7273, Santa, USA), CEBPα (sc-365318, Santa, USA), Ang-1 (23302-1-AP, Sanying, China) and VEGF (66828-1-Ig, Sanying, China) overnight at 4 °C.

Techniques: Western Blot, Labeling