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Figure 2 | Breast Cancer Research

Figure 2

From: A mammosphere formation RNAi screen reveals that ATG4A promotes a breast cancer stem-like phenotype

Figure 2

Negative selection mammosphere formation shRNAi screen. (A) Schematic, illustrating layout of pooled RNAi screen. Cells were transduced with the lentiviral DECIPHER RNAi library pool at low multiplicity of infection and cultured for fourteen days under adherent or suspension conditions, respectively. At the end of the screen, adherent cells and mammospheres >40 μm were harvested. Using next-generation sequencing of barcodes, the abundance of each shRNA expression construct in the pool was determined at the beginning of the screen (baseline) and from adherently cultured cells and mammospheres at fourteen days post seeding. Colour scheme: red: toxic shRNA; yellow: shRNA differentially inhibiting sphere formation; green: shRNA differentially activating sphere formation. (B) Barcode ranking. Shown are log2 (tsphere/baseline) read-count ratios for all 27,500 shRNA expression constructs analysed in the screen. (C) Selected candidate genes involved in Janus kinase (Jak)-signal transducers and activators of transcription (STAT) Jak-STAT signalling.

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