RNA-seq of U2OS cells treated with control or LSm4-targeting siRNAs, alone or with expression of either LSm4-WT or LSm4-del69-92
RNA was collected from 6-well plates using TRIzol reagent (Ambion) according to the manufacturer's protocol. Total RNA was isolated using TRIzol reagent (Ambion) according to the manufacturer's instructions. In brief, chloroform was added and after vortexing, sample was centrifuged for 15 minutes at 12,000 x g, 4 degrees. Upper phase was collected, RNA was precipitated in -20 degrees using glyco blue and isopropanol, and the sample was then centrifuged for 15 minutes at 12,000 x g, 4 degrees. Then, samples were washed with ice-cold 70% ethanol and centrifuged for 5 minutes at 7,500 x g, 4 degrees. Pellet was dried and reconstituted in nuclease-free water on ice. RNA sequencing libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB, cat no. E7760).