Celltitre glo 2

Cells were seeded into a white-walled, glass-bottomed 384-well plate (Nunc) at a density of 1 × 103 cells per well. The cells were treated with escalating doses of erlotinib, osimertinib or CX-4945 24 h following seeding over a period of 72 h. Cell viability was determined using CellTitre-Glo 2.0 (Promega Corporation) according to the manufacturer’s instructions. Luminescence was scanned and analysed on the FLUOstar Omega Microplate Reader (BMG Labtech, Mornington, Australia). Data were normalised to untreated controls and dose–response curves and drug potency values generated using GraphPad Prism V9 software.

Cells were seeded into a white-walled, glass-bottom 384-well plate (Nunc) at a density of 1 × 10³ cells per well. 24 h following seeding, the cells were treated with escalating concentrations of 5-FU alone or for the sequential combination approach, a low and high concentration of AZD1152 was added 24 h following treatment with 5-FU. Cell viability was determined using CellTitre-Glo 2.0 (Promega Corporation), according to the manufacturer’s instructions, 72 h following commencement of drug treatments. Luminescence was measured and analysed on the FLUOstar Omega Microplate Reader (BMG Labtech). Data was normalised to untreated controls, and dose response curves and drug potency values generated using GraphPad Prism V9 software.

The viability response of cells was assessed using CellTitre-Glo 2.0 (Promega, Cat#G9242), and luminescence was quantitated by a plate reader. Cells were plated in triplicate in a flat bottom 96-well plate (Corning, Cat#3903) at starting concentrations of either 2 × 10⁴ or 2 × 10⁵, depending on cell doubling time. Drug treatments were assessed between 24–144 h. All measurements were performed based on the manufacturer’s protocol.