Nature research paper: CCNE1 amplification is synthetic lethal with PKMYT1 kinase inhibition
PCNA-cb-TagRFP expressing cells were maintained at 37 °C and 5% CO2 while deconvolution wide-field microscopy was performed using the DeltaVision Elite system equipped with an NA 0.75 20× UPlanSApo objective and an sCMOS 2,048 × 2,048 camera . Each field was acquired every 10 min over 23 h with a-step of 2 μm through the entire cell and deconvolved using softWoRx . Maximum intensity projections are shown .
Cells were analysed at the LTRI flow cytometry facility on a Fortessa X-20 using FACSDIVA v8.0.1 with at least 9,000 events collected and analysed using FlowJo v10.Cell pellets were resuspended in 250 μl EBN buffer , 15 mM MgClovalbumin , 1× protease inhibitor cocktail pH 7.3) and incubated on ice for 5 min.