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. 2021 Sep 23;116:105362. doi: 10.1016/j.bioorg.2021.105362

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© 2021 Published by Elsevier Inc.

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Fig. 3 — Validation of the FP assay. A) FP readings of 10 nM of HR2P-FL; 10 nM of HR2P-FL with 1 μM of NusA, 5-HB, or NusA-5HB. B) Determination of the six-helix bundle formation between HR2P-FL and 5-HB/NusA-5HB by fluorescence N-PAGE. Lane 1: 10 μM HR2P-FL; Lane 2: 10 μM HR2P-FL + 10 μM HR1P; Lane 3: 10 μM HR2P-FL + 10 μM 5-HB; Lane 4: 10 μM HR2P-FL + 10 μM NusA-5HB. C) Saturation binding FP experiments. The FP readings were converted to fraction bounded (f), and then plotted against the NusA-5HB concentration to get K_d = 3.2 nM. All experiments were performed in duplicate.