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. 2021 May 6;7(6):980–989. doi: 10.1021/acscentsci.0c01592

Figure 6.

Figure 6

Compound 9a directly binds NCOA4383–522 and inhibits the NCOA4383–522–FTH1 interaction. (A) Specific binding of 9a to NCOA4383–522. The recombinant NCOA4383–522 or FTH1 protein was pretreated with 9a or DMSO for 1 h and then incubated with increasing concentrations of Biotin-14d (precoupled with streptavidin magnetic beads) for 1 h. The proteins bound to the beads were detected by silver staining and Western blotting. (B) SPR sensorgrams for the binding of 9a and the inactive compound 9h to NCOA4383–522. (C) Schematic of the ELISA assay used to detect the NCOA4383–522–FTH1 interaction and its disruption by 9a. (D) Dose–response relationship for the inhibition of the NCOA4383–522–FTH1 interaction by 9a. The indicated concentration of 9a was preincubated with NCOA4383–522, and the binding of NCOA4383–522 to FTH1 was monitored by ELISA. Data shown represent the mean ± SEM from three independent experiments.