Human cell cytokine release assay
Tecan
Frozen PBMCs were thawed into RPMI with heat-inactivated 10% FBS/1% penicillin–streptomycin and the same day plated into 96-well flat-bottom plates at 200,000 cells per well in 190 μl of media. KT-474, PF-06550833 and DMSO controls were prepared in duplicate for each donor. All cells were dosed using the Tecan automated liquid handler, followed by incubation at 37 °C and 5% CO