Furamidine

H1975 and H3255 cells were seeded at 5,500 cells per well in a 96-well plate to adhere and grow for 24 hours. Cells were then transfected with siRNA using Dharmafect-2 (GE Life Sciences) for 24 hours, followed by 10 μM Erlotinib for 24 hours. For treatment with a PRMT-1 inhibitor Furamidine (Tocris Bioscience), cells were incubated at 37°C in media containing 20–25μM Furamidine for 24 hours, followed by Erlotinib for an additional 24 hours. Finally, MTT cell viability assays were performed as described previously[2 (link)].

Cells were seeded in 96-well plates at 2,000–10,000 cells per well, depending on the cell line. For single-agent dose response assays, furamidine (Tocris, #5202) or MS023 (Tocris, #5713) was added at the indicated concentrations using a D300e Digital Dispenser (Tecan) or by manual serial dilution, with DMSO treatment as a negative control. After 7 days, cell viability was measured with the CellTiter-Glo Luminescent Cell Viability Assay (Promega, #G7571) and normalized to DMSO control wells. For experiments evaluating combined AR and PRMT1 inhibition, cells were treated with a combination of furamidine (Tocris) and enzalutamide (Selleck) doses as indicated. After 7 days, cell viability was measured by CellTiter-Glo and normalized to DMSO wells. Drug synergy was evaluated with a Bliss independence model using Combenefit v2.021 (Di Veroli et al., 2016 (link)).