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. Author manuscript; available in PMC: 2016 Jan 15.
Published in final edited form as: J Med Chem. 2015 Aug 28;58(17):6938–6959. doi: 10.1021/acs.jmedchem.5b00801

Figure 6.

Figure 6

FRα and FRβ binding assays for assorted antifolates. Data are shown for the relative binding affinities of the unlabeled ligands with FRα-expressing RT16 and FRβ-expressing D4 CHO cells. Relative binding affinities for assorted folate/antifolate substrates were determined over a range of ligand concentrations and were calculated as the inverse molar ratios of unlabeled ligands required to inhibit [3Hfolic acid binding by 50%. By definition, the relative affinity of folic acid is 1. Results are presented as mean values plus/minus standard errors from 4–10 experiments. The detailed methods are provided in the Experimental Procedures. Abbreviations: Cpd, compound; FA, folic acid.