Identification of human gene LOC107984012 interacting proteins in AC16 cardiomyocytes by pull-down assays coupled to MS

In this study, pull down assays combined with mass spectrometry analysis were performed in AC16 cardiomyocytes transfected with control plasmid (OE-Vector) or LOC107984012-overexpression plasmid to identify the repertoire of LOC107984012 interacting proteins. Pierce™ Magnetic RNA-Protein Pull-Down Kit (Thermo Fisher) provides reagents to efficiently enrich RNA Binding Proteins. Sense or antisense of LOC107984012 were in vitro transcription using the T7 RNA transcription system (Large Scale RNA ProductionSystem-T7, Promega) and biotin-labeled with the Pierce™ RNA 3’ End Desthiobiotinylation Kit (Thermo Scientific). RNA was then purified with QIAquick PCR Purification Kit (Qiagen). 30 μg of whole-cell protein lysates were incubated with purified biotinylated transcripts for 1 h at 4°C with rotation, then they were eluted for mass spectrometry identification.