plasmid encoding human mpa-gfp-cd81-10 #57124  (Addgene inc)

Journal: Journal of Hematology & Oncology

Article Title: Tumor stem-like cell-derived exosomal RNAs prime neutrophils for facilitating tumorigenesis of colon cancer

doi: 10.1186/s13045-019-0699-4

Figure Lengend Snippet: SDCSC exosomes are transported to bone marrows and increase neutrophil population in bone marrow. a Representative images of exosomes captured with TEM. Scale = 100 nm. b The size distribution of exosomes by NTA. c Western blots of exosome markers in cells. P, parental cells; S, SDCSCs; WCL, whole cell lysate. d Histograms showing relative exosome secretion in CRC cells. * P < .05. e The representative images for transferring of PKH26-labeled exosomes. Scale bar = 10 μm. f The relative luciferase activity of purified exosomes. g The relative luciferase activity of CT26 cells receiving CT26-SDCSC-released exosomes. GFP-Luc-SDCSC exosome, exosomes from GFP-luciferase-expressing SDCSCs. * P < .05. h The relative luciferase activity in bone marrow cells of normal mice ( N = 5), mice with tumors grown from GFP-Luc parental cells ( N = 7), and mice with GFP-Luc SDCSC-derived tumors ( N = 7). The data represent mean ± SEM. * P < .05. i Western blot of CD81 and FLAG from 10 μg of exosome proteins purified from ectopically FLAG-CD81-expressing SDCSCs and GFP-expressing control cells. j Left: representative images of FLAG-FITC signals in cells. Right: quantification of FLAG-FITC positive cells. *** P < .001

Article Snippet: The CD81 fragment of mPA-GFP-CD81-10 (Plasmid #57124, Addgene, Cambridge, MA, USA) was employed to generate pCDH-FLAG-CD81.

Techniques: Western Blot, Transferring, Labeling, Luciferase, Activity Assay, Purification, Expressing, Derivative Assay, Control