Sequencing of RNA fragments immunoprecipitated by anti-Flag in CASC3 or MTDH overexpression breast cancer cells
RNA immunoprecipitated sequencings were conducted from CASC3 or MTDH overexpression breast cancer cells Overall design: Breast cancer cells MD-MBA-231 was transfected with pEZ-93-CASC3 or pEZ-93-MTDH plasmids, which contain Flag sequence and over-express CASC3 or MTDH in cells. Bound complexes were pulled-down using an antibody against Flag. Then RNA immunoprecipitation (RIP) assays were performed with a Magna RIP Kit (Millipore). The RNA samples from RBP-OV and control cells were prepared and analyzed by the Agilent 2100 system. The library was conducted by the KAPA Stranded RNA-Seq Library Prep Kit (Illumina, USA), and sequenced on the Illumina NovaSeq 6000 using the NovaSeq 6000 S4 Reagent Kit (Illumina).