cd63 gfp vector  (Addgene inc)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Exosomal transfer of miR-15b-3p enhances tumorigenesis and malignant transformation through the DYNLT1/Caspase-3/Caspase-9 signaling pathway in gastric cancer

doi: 10.1186/s13046-019-1511-6

Figure Lengend Snippet: Exo-miR-15b-3p high expression in GC patient serum as a GC diagnosis and prognosis biomarker. a Representative electron microscopy micrographs of SGC-7901, BGC-823 and GES-1 cell conditioning medium secreted exosomes, as well as control and GC patient serum secreted exosomes. Scale bar, 100 nm. b Nano Sight particle-tracking analysis to determine exosome size distribution and number. c Levels of exosomal markers, TSG101, CD63 and CD9, of cells and serum derived exosomes determined using western blotting analysis. d Relative expressions of Exo-miR-15b-3p in conditioned medium of BGC-823, SGC-7901 and GES-1 cells. e Relative exosomal miR-15b-3p levels in GC patients and non-GC normal volunteer ( n = 108) serum. f The sensitivity and specificity of serum Exo-miR-15b-3p for GC prediction was evaluated through Receiver-operating characteristic (ROC) curve analysis. g The correlation between serum Exo-miR-15b-3p expression and overall survival of the 108 GC patients determined using Kaplan–Meier analysis. The cutoff used was the median. Mean ± SEM of the results are presented

Article Snippet: Genechem Inc. (China) constructed luciferase-labelled lentivirus vectors carrying miR-15b-3p (Lv-miR-15b-3p)/negative control (Lv-NC), miR-15b-3p inhibitor (Lv-inhibitor)/negative control (Lv-inNC) and GFP-labelled lentivirus vectors containing CD63 (GFP-Lv-CD63) were used.

Techniques: Expressing, Biomarker Discovery, Electron Microscopy, Control, Derivative Assay, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Exosomal transfer of miR-15b-3p enhances tumorigenesis and malignant transformation through the DYNLT1/Caspase-3/Caspase-9 signaling pathway in gastric cancer

doi: 10.1186/s13046-019-1511-6

Figure Lengend Snippet: Exosome-mediated miRNA transport between cells. a Internalization of PKH26-labelled exosomes (red) in GES-1 and SGC-7901 cells were observed through confocal microscopy. Fluorescein phalloidin-FITC (green) was used to stain F-actin, while DAPI (blue) was used to stain nuclei. Scale bar, 20 μm. b Exosomes (green) isolated from BGC-823 cell conditioning medium labeled with GFP-Lv-CD63 (green) and transfected with Cy3-miR-15b-3p (red) were co-cultured with SGC-7901 and GES-1 cells for 24 h, and the fluorescent signals were detected using confocal microscopy. Nuclei are stained blue (DAPI). Scale bar, 20 μm. c The efficiency of exosomes in delivering miR-15b-3p to GES-1 and SGC-7901 cells was analyzed using RT-PCR. Mean ± SEM of the results are presented

Article Snippet: Genechem Inc. (China) constructed luciferase-labelled lentivirus vectors carrying miR-15b-3p (Lv-miR-15b-3p)/negative control (Lv-NC), miR-15b-3p inhibitor (Lv-inhibitor)/negative control (Lv-inNC) and GFP-labelled lentivirus vectors containing CD63 (GFP-Lv-CD63) were used.

Techniques: Confocal Microscopy, Staining, Isolation, Labeling, Transfection, Cell Culture, Reverse Transcription Polymerase Chain Reaction