Development and application of high-throughput screens for the discovery of compounds that disrupt ErbB4 signaling: Candidate cancer therapeutics
Fig 2
Validation of semi-automated processes for the stimulation of ErbB4-dependent cellular proliferation and detection of the inhibition of agonist-induced ErbB4-dependent cellular proliferation.
(a) BaF3/EGFR+ErbB4 cells were stimulated with increasing concentrations of NRG1β in four independent trials using a semi-automated protocol. A semi-automated MTT assay was used to analyze cell proliferation 120 hours post-stimulation. Curves were fit to the data using GraphPad Prism to determine the EC50 of NRG1β. The Z-factor for ErbB4-dependent cell proliferation stimulated by 0.3 nM NRG1β was calculated using the means and standard deviations of the positive (0.3 nM NRG1β) and negative (NRG1β-diluent) controls. (b) BaF3/EGFR+ErbB4 cells were treated with increasing concentrations of gefitinib in the presence of 0.3 nM NRG1β in four independent trials using a semi-automated protocol. A semi-automated MTT assay was used to analyze cell proliferation 120 hours post-stimulation. Curves were fit to the data using GraphPad Prism to determine the IC50 of gefitinib against NRG1β.