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. 2015 Jun 11;24(7):1129–1146. doi: 10.1002/pro.2701

Figure 4.

Figure 4

SPR measurements of Rev-erbα LBD interactions. SPR response (expressed as RU) of different modulators on Rev-erbα LBD (313 nM) binding to biotinylated-NCoRID1 immobilized on a streptavidin chip. (A) Competition of NCoRID1 binding to Rev-erbα LBD by addition of SmrtID1 (Rev-erb alone: yellow brown; 0.1—red, 0.3—green, 1.0—blue, 3.0—pink, and 10—cyan µM SmrtID1); (B) effect of the ligand SD7 (25 µM) on NCoRID1/Rev-erbα LBD interaction (no ligand—blue; 25 µM SD7—red); (C) effect of the ligand SGN (12.5 µM) on NCoR/Rev-erbα LBD interactions (no ligand—blue, 12.5 µM SGN—red) ; (C) Dose–effect response of the ligand SGN on NCoRID1/Rev-erbα LBD interactions (no ligand—red, 3.12 µM SGN—green; 6.25 µM—blue; 12.5 µM—pink; 25 µM—cyan; 50 µM—yellow brown; 100 µM gray). EC50 determination in the insert (EC50 = 20 µM). The percentage of activation of Rev-erbα LBD (313 nM) binding to immobilized NCoRD1 in the presence of SGN is reported in the insert and this allowed EC50 determination (EC50 = 20 µM). All experiments were performed in duplicate. Experiments were analyzed in PRISM® software using a one phase exponential association fitting method.