Esophageal Adenocarcinoma Methods and Protocols

257 Alfred K. Lam (ed.), Esophageal Adenocarcinoma: Methods and Protocols, Methods in Molecular Biology, vol. 1756, https://doi. ...

258 during the pathogenesis of esophageal adenocarcinoma. They could be markers for prediction of progression of Barrett’s esoph ...

259 2 Materials Total RNA extraction kit. Chloroform. 1.5 μL tube. 2.1 Isolation of Total RNA Fig. 1 Schematic presentation of ...

260 RNase-free water. 100% ethanol. Centrifuges. RNase-free, sterile-filtered water. miRNA isolation kit. 2× hybridization buff ...

261 Standard thermal cycler. Centrifuge with plate holders. Microcentrifuge. Vortex. Polypropylene tube. Reagent tubes with cap ...

262 infer relative changes in expression between samples. Microarrays are useful as they are able to screen large number of miRN ...

263 Denature by incubating at 95 °C for 2 min and immediately cooling down on ice (see Note 8). Put the backing slide in the ch ...

264 lengthening the molecule by incorporating a poly(A) tail or stem- loop structure [ 31 ]. Consequently, cDNA can be used in q ...

265 Mix and centrifuge. Dispense 1.0 μL (or alternative according to the kit manual) of 20× MicroRNA Assay mix into the corresp ...

266 is distributed equally across the slide. Take care to not touch the slide! Avoid dropping the array slide onto the gasket s ...

267 roles and current perspectives. Exp Mol Pathol 96:98–107. https://doi.org/10.1016/j. yexmp.2013.12.004 Maroof H, Salajegheh ...

268 Leshkowitz D, Horn-Saban S, Parmet Y, Feldmesser E (2013) Differences in microRNA detection levels are technology and seque ...

269 Alfred K. Lam (ed.), Esophageal Adenocarcinoma: Methods and Protocols, Methods in Molecular Biology, vol. 1756, https://doi. ...

270 the successful delivery of the double-stranded small RNA molecules to the target cells. There are mainly two types of short ...

271 MET is a receptor tyrosine kinase, act as a proto-oncogene and promotes the different stages of pathogenesis of esophageal a ...

Fig. 1 Delivery and mechanism of RNAi via shRNA lentiviral particles in esophageal adenocarcinoma cells. Lentiviral particles de ...

273 using lentiviral particles expressing MET shRNA. Subsequently, we select the stable clones of cells with antibiotic. Finally ...

274 Cell lysis radioimmunoprecipitation assay (RIPA) buffer. 150 mM sodium chloride, 1.0% NP-40 or Triton X-100, 0.5% sodium de ...

275 Count the cells and plate 0.5–2.0 × 105 cells in a 12-well plate 24 h prior to viral transfection. Add 1 mL complete media ...

276 Prepare gel for electrophoretic separation of the protein mix- ture. First, preparation of separation gel (10%): Add 4.1 mL ...