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. 2023 Mar 21;66(7):5208–5222. doi: 10.1021/acs.jmedchem.3c00151

Figure 4.

Figure 4

NanoBRET saturation binding curves measured in HEK293G NLuc-CXCR4 cell membranes for (a) 10, (b) 11, (c) 18a, (d) 18b, and (e) 24. Raw BRET ratio was determined by the ratio of fluorescence/luminescence emissions. Measurements were made in the absence (total binding) and presence of 10 μM AMD3100. For those curves (a,c,e) where the non-specific binding curves obtained in the presence of 10 μM AMD3100 were not linear, pKd values were estimated by fitting the total binding curves to a saturable plus linear component. Data points represent mean ± S.D. of triplicate determinations of a single experiment. These single experiments are representative of 3–5 separate experiments per fluorescent ligand. Where not seen, error bars are within the symbol. White opaque-bottom 96-well plates were used for compounds 10 and 11 (a,b), whereas white clear-bottom 96-well plates were used for compounds 18a, 18b, and 24 (c–e).