RNA-seq of 293T cell line
293T/MPL cells were treated with THPO (100 ng/L) for 30min or control in three replicates. Cells were washed with ice-cold PBS and crosslinked with UV-light at 254nm. Nuclear extracts were subjected to immunprecipitation using an anti-HNRNPA1 antibody (clone 4B10) or a monoclonal murine IgG2A kappa isotype control antibody using protein-A beads (Promega). RNA of the washed immunoprecipitates and unmanipulated input was extracted using TRIZol, ERCC spike in RNA added and libraries were generated using the SMART-Seq Total RNA Pico Input with UMIs (stranded) Kit (Takara)