GSM8118596
To study the influence of PABPC1 SUMOylation on mRNA stability, we performed 4sU pulse-chase sequencing in H1299 cells under normal or stress condition to detect RNA stability. Briefly, H1299 cells stably expressing PABPC1-WT or PABPC1-K512R were labeled with 200 mM 4-thio-uridine for 2 h, after which medium was replaced with fresh medium with or without 500 µM sodium arsenite. The total RNA was extracted at indicated time point, then 4sU labeled RNAs were converted to biotinylated RNAs using HPDP-Biotin and isolated using DynabeadsTM MyOneTM Streptavidin C1. 4sU-labeled RNA was used for removing the rRNAs using Ribo-Zero rRNA Removal Kits (Illumina, USA) following the manufacturer's instructions. RNA libraries were constructed by using rRNA-depleted RNAs with TruSeq Stranded Total RNA Library Prep Kit (Illumina, USA) according to the manufacturer's instructions. mRNA-Seq and LncRNA-Seq