GSM5352228: Set-1_shTP53-RB1-2; Homo sapiens; RNA-Seq
For RNAseq samples: LNCaP/AR cells transduced with different CRISPR constructs were treated with enzalutamide or vehicle for 6 days before the total RNA was extracted using Trizol. For scRNAseq samples: Based on FACS analysis, single cells were sorted into 1.5 ml tubes (Eppendorf) and counted manually under the microscope. The concentration of single cell suspensions was adjusted to 900-1100 cells/μl. Cells were loaded between 10,000 and 17,000 cells/chip position using the Chromium Single cell 5' Library, Gel Bead & Multiplex Kit and Chip Kit (10x Genomics, V1 barcoding chemistry). For RNAseq samples: RNA-Seq libraries were prepared using the Illumina TruSeq stranded mRNA kit, with 10 cycles of PCR amplification, starting from 500 ng of total RNA. For scRNAseq samples: Single-cell gene expression libraries were generated according to the manufacturer's instructions. All the subsequent steps were performed following the standard manufacturer's protocols. For RNAseq samples: RNA-Seq. For scRNAseq samples: Single-cell RNA-seq were performed by the 10x Genomic single cell 5'library platform.