Fig. 5

FH is identified as the direct target of rutaecarpine. A. Schematic for DARTS assay to identify the target of small molecules. B. SDS-PAGE for the DARTS assay, highlighting and purifying the potential targeting proteins specific to 3D and 2D culture for further MS analysis. C. KEGG analysis for the potential targeting proteins of rutaecarpine showing enrichment in carbon metabolism. D. Western-blotting showing a dose dependent function of rutaecarpine in preventing the degradation of FH by pronase in cell lysates from MDA-MB-231 cells in spheroids. E. Western-blotting showing a dose dependent function of rutaecarpine in preventing the degradation of FH purified from bacteria. F. Western-blotting showing that rutaecarpine has no effect in regulating the protein level of FH in MDA-MB-231 cells from 2D and 3D cultures