A total of 800 NMuMG or SNU-1 cells; 2,000 MCF7, MDA-MB-231, KATO-III, SNU-16, or SUM52PE cells; 3,000 MFM-223 or NCI-H716 cells; or 4,000 MDA-MB-134-VI cells per well were seeded in 96-well plates using DMEM/F-12 supplemented with penicillin–streptomycin and 10% FBS for human cell lines or 3% FBS for NMuMG. After 24 h, cells were treated with FGFRi for 4 days using vehicle (DMSO), AZD4547 (AstraZeneca), or pemigatinib (HY-109099), BGJ398 (HY-13311) or debio-1347 (HY-19957, all MedChemExpress) with a range of 0.1 nM to 100 μM. Usage of AZD4547, pemigatinib, BGJ398 and debio-1347 was previously described63 (link)–66 (link). Cell viability was assayed using CellTiter-Blue Reagent (G808A, Promega) for 4 h and subsequently measuring fluorescence on the Infinite M Plex plate reader operated using the Tecan i-control software. Drug-response curves were modelled using [inhibitor] versus response with variable slope (four parameters) and least-squares regression in Prism (v.9.3.1, GraphPad Software).
Zingg D., Bhin J., Yemelyanenko J., Kas S.M., Rolfs F., Lutz C., Lee J.K., Klarenbeek S., Silverman I.M., Annunziato S., Chan C.S., Piersma S.R., Eijkman T., Badoux M., Gogola E., Siteur B., Sprengers J., de Klein B., de Goeij-de Haas R.R., Riedlinger G.M., Ke H., Madison R., Drenth A.P., van der Burg E., Schut E., Henneman L., van Miltenburg M.H., Proost N., Zhen H., Wientjens E., de Bruijn R., de Ruiter J.R., Boon U., de Korte-Grimmerink R., van Gerwen B., Féliz L., Abou-Alfa G.K., Ross J.S., van de Ven M., Rottenberg S., Cuppen E., Chessex A.V., Ali S.M., Burn T.C., Jimenez C.R., Ganesan S., Wessels L.F, & Jonkers J. (2022). Truncated FGFR2 is a clinically actionable oncogene in multiple cancers. Nature, 608(7923), 609-617.
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