RNA-Seq of U87 cell line: shNC
A total amount of 1g qualified RNA per sample was used as input material for the library preparation. The sequencing libraries were generated using the VAHTS mRNA-seq v2 Library Prep Kit for Illumina (Vazyme, NR601) following manufacturers recommendations. Firstly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was performed using divalent cations under elevated temperature in Vazyme Frag/Prime Buffer. The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers. Second strand cDNA synthesis was subsequently performed using buffer, dNTPs, DNA polymerase I and RNase H. Then, the cDNA fragments were end repaired with the addition of a single A base at the 3'-end of each strand, ligated with the special sequencing adapters (Vazyme, N803) subsequently. The products were purified and size selected with VAHTSTM DNA Clean Beads (Vazyme, N411) in order to get appropriate size for sequencing. Following amplification, cDNA was purified and subjected to library construction for the paired-end 150 bp (PE150) sequencing on Illumina HiSeq X Ten platform.