ELAVL1 (NM_001419) Human Recombinant Protein

Snippet: release bound protein from the beads. Samples were processed for Western blot analysis as above. Electrophoretic mobility-shift analysis Recombinant HuR (4.5 - 120 nM), obtained from OriGene Technologies (cat. no. TP301562), was incubated ... Cite SaveSaving

Snippet: HUVECs were seeded 1 d before transfection at 3.5 × 105 cells per well and transfected with 5 µg of either pLenti-ADAR1 (Origene) or an empty pLenti control (Origene) (both plasmids bear the SV40 origin of replication (SV40 ori)) using Lipofectamine 2000 transfection reagent (Invitrogen), according to the manufacturer’s instructions. After 24–48 h, the transfected cells were subjected to a 3D sprouting assay or to gene expression and RNA editing studies, as described below. 3 × 106 HeLa cells (grown in Dulbecco’s modified Eagle’s medium (DMEM;Lonza) with 10% FBS supplied with antibiotics) that were seeded onto a T75 flask (Greiner) 1 d before transfection were transfected with 5 µg of pCMV6-XL-ADAR1 (Origene) using GeneJuice transfection reagent (Novagen) and incubated for 24 h before the in vitro RNA editing assay, which is described below