Nature research paper: RBFOX2 modulates a metastatic signature of alternative splicing in pancreatic cancer
Lentiviruses were produced by co-transfection of HEK293T cells with psPax2 , pMD2.G and either Flag SFFV2 ΔRRM RBFOX2-puro or specific sgRNAs LentiCRISPRV2, using FuGENE-HD and OptiMEM . One day after transfection, the medium was replaced, and 48 h after transfection, viruses were collected and filtered through a 0.45 µm membrane. BxPc3, X50, and X252 cells were infected with the viruses with the addition of polybrene . Selection with puromycin was initiated 2 days after infection.
Cell density was determined every 24 h by absorbance of the methylene blue dye at 655 nm, measured on a plate reader according to the manufacturer’s instructions. For MBQ167 treatment, cells were treated with either 0.05 µM MBQ167 or DMSO for 24 h after seeding.
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