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Wild-type (WT) 3′-UTR fragments of circUSP9X and SRCIN1 containing predicted miR-148b-3p binding sites were amplified and cloned into the pmirGLO Dual-Luciferase Expression Vector (Promega, Madison, WI, USA) to construct reporter vectors SRCIN1-WT and circUSP9X-WT. The putative binding ...
MTS method was conducted to assess the effects of drugs on cell viability using CellTiter 96 AQueous One Solution Cell Proliferation Assay (G3580; Promega, Madison, WI) according to the manufacturer’s protocol. After incubating for 3 h, we further transferred the media with MTS to a 96-...
For each reac- tion in a 96-well plate, 100 ng of protein was used, and the ELISA was performed according to the manufacturer's protocol (Promega, Madison, WI, USA). A standard curve was included in each experiment. Western blotting. Total protein was extracted from cells using ...
A total of 2 μg of high quality RNA was processed directly to cDNA with the reverse transcription kit (Promega, Madison, WI, USA), following the manufacturer's instructions. The PCR procedure used for the detection of human cIAP1, EGFR, Integrin β-5, uPAR, cMet and β-actin was ...
The samples were then incubated with 1 μg of modified trypsin (Promega, Madison, WI, USA) for 16 hours. Enrichment of phosphopeptides An immobilized metal ion affinity chromatography (IMAC) column (5 cm × 500 μm i.d., PEEK tubing, which is an attached component of spin column produced...
https://ror.org/01y2jtd41grid.14003.360000 0001 2167 3675Interventional Radiology, Department of RadiologyUniversity of Wisconsin-Madison 600 Highland Avenue 53792 Madison WI USAJohn Swietlikhttps://ror.org/01y2jtd41grid.14003.360000 0001 2167 3675Interventional Radiology, Department of RadiologyUniversity...
Phosphorothioate antisense oligonucleotides (denoted with the prefix “as”) used in the present study were designed on the known cloned RYR sequences deposited in the GenBank™ sequence data base with Lasergene software (DNASTAR, Madison, WI). Sequences of all three RYR cDNAs were aligned wit...
; pT7Blue T-vector (NOVAGEN, Inc., WI); pGEM-3Z/4Z (PROMEGA Inc., Madison, Wis.), and the like vectors, such as is described herein. Exemplary vectors for expression using Agrobacterium tumefaciens-mediated plant transformation include pBin 19 (CLONETECH), Frisch et al, Plant Mol. Biol...