Extended Data Fig. 2: Recurrent genetic alterations associated with bladder and prostate cancer encoded in barcoded lentiviral libraries.

(a) Tables showing gain-of-function and loss-of-function genetic alterations associated with bladder and prostate cancer selected for representation in cancer-specific barcoded lentiviral libraries. (b) Schematics of barcoded lentiviral vectors expressing open reading frames (ORF) or short-hairpin RNA (shRNA). LTR = long terminal repeat; BC = barcode; UBC = Ubiquitin C; CMV = cytomegalovirus; GFP = green fluorescent protein; WHV8 = Woodchuck hepatitis virus 8 post-transcriptional regulatory element. (c) Plot showing relative expression of target genes as determined by quantitative polymerase chain reaction (qPCR) in 3T3 cells 72 hours after lentiviral transduction with pLKO.1-TRC control or pLKO.1 expressing select shRNA previously screened and selected for inclusion in the barcoded lentiviral libraries based on the extent of gene knockdown. qPCR reactions were performed on four biologically independent replicates. Statistical analysis was performed by two-tailed, unpaired t-test with p-values shown. (d) Plot showing relative overexpression of gain-of-function genes as determined by qPCR in 3T3 cells 72 hours after lentiviral transduction with barcoded ORF vectors. qPCR reactions were performed on three biologically independent replicates. Statistical analysis was performed by two-tailed, unpaired t-test with p-values shown.