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Quantitative real-time PCR Quantitative real-time PCR was performed using a total reaction volume of 10 l, containing 2 l of diluted cDNA, SsoAdvanced Universal SYBR Green Supermix (1x) (Bio-Rad), and a final concentration of 0.25 M or 0.5 M of forward and reverse primers (designed using Primer-BLAST, Table S5), with a CFX Connect Real-Time Cycler and CFX manager (Bio-Rad)

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Cancer Cell 10:375-388, 2006

MDA-MB-231, MDA-MB-468 and MDA-MB-436 cells were seeded in 6-well plates with 10 6 /10 5 cells/well containing 3 mL media and the plates were incubated at 37 C overnight

The samples were analysed as a single batch to reduce batch-to-batch variation and randomised to account for LC-MS system drift over time