library_construction_protocol: B67 GSCs were infected with MISSION TRC lentiviruses with shRNA targeting human TRIM26 (sh4050, sh4051) or with control (sh002) and 5 days later harvested for total RNA Total RNA was isolated using the RNeasy Plus Mini Kit (Qiagen). Ribosomal RNA was removed by a hybridization method using Ribo-ZERO kits (Illumina).  mRNA was fragmented and reverse transcribed to yield double-stranded cDNA. cDNA was blunt ended, had an A base added to the 3â€² ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 12 cycles using primers incorporating unique index tags.