RNA-seq of human breast cancer cell line MCF7, T47D and CAMA-1 treated that are naïve and resistant to capivasertib and have been treated with acute capivasertib, continuous capivasertib and DMSO
RNA-seq of human breast cancer cell line MCF7, T47D and CAMA-1 treated that are naïve and resistant to capivasertib and have been treated with acute capivasertib, continuous capivasertib and DMSO
Cells were cultured for 6 months with increasing doses of capivasertib up to 10μM. Once generated, CapiR cells were routinely cultured in RPMI media (Gibco #11835-063) supplemented with 10% Fetal Calf Serum (FCS), 1% Glutamax, 10μM capivasertib and incubated at 37°C, 5% CO2. RNA was extracted from the treated cells using the AllPrep RNA Mini Kit (Qiagen, 80504). RNA-Seq was performed with poly(A) selection using standard methods ibraries were quantified with the D1000 ScreenTape assay (Agilent; 5067-5582, 5067-5583) on the Tapestation 4200 system (Agilent) and subsequently pooled equimolar. The final pooled library was loaded onto two lanes of a NovaSeq X Series 25B flow cell (Illumina; 20104706) and run at a paired end 150 base pair configuration (300 cycles) with 1% PhiX spiked in on a NovaSeq X Plus (Illumina).