Figure 1. A genetic screen for drivers of anchorage-independent growth in human mammary epithelial cells.
(A) A schematic of the screen. TL-HMECs were transduced with the ORFeome library (8000 CMV promoter-driven human open reading frames [ORFs] in a retroviral vector) at a multiplicity of infection (MOI) of 0.2 and plated into semi-solid medium. Macroscopic colonies were isolated, individually expanded, and the identities of ORF inserts were determined by sequencing. (B) ORFs recovered from two independent screen replicates were individually transduced into TL-HMECs and plated into semi-solid medium. Colonies were counted and colony numbers were normalized to an empty vector-transduced sample. Asterisks denote strongly validated ORFs that localize to focal amplification peaks in at least one tumor subtype. Assays were performed in triplicate (error bars ± SD).