Fig. 2

Rutaecarpine induces luminal differentiation of TNBC cells in 3D culture. A. Spheroid formation from MDA-MB-231 and BT549 cells with DMSO or rutaecarpine treatment. B. Quantification of circularity and Feret’s diameter for spheroids formed by MDA-MB-231 and BT549 cells. Unpaired Student’s t-test; **** p < 0.0001. C. IF staining of KRT8 in DMSO or rutaecarpine-treated MDA-MB-231 and BT549 spheroids. D. Quantification of KRT8 MFI of the DMSO or rutaecarpine-treated MDA-MB-231 and BT549 spheroids. Unpaired Student’s t-test; ** p < 0.01; **** p < 0.0001. E. qPCR of luminal marker genes in DMSO or rutaecarpine-treated MDA-MB-231 spheroids. Unpaired Student’s t-test; *** p < 0.001. F. qPCR of luminal marker genes in DMSO or rutaecarpine-treated BT549 spheroids. Unpaired Student’s t-test; ** p < 0.01; *** p < 0.001; **** p < 0.0001. G. GSEA analysis showing significant enrichment of apoptosis genes in rutaecarpine-treated MDA-MB-231 cells in 3D spheroids. H. Apoptosis analysis for the matured MCF-10A spheroids with PI staining. I. Apoptosis analysis for DMSO or rutaecarpine-treated MDA-MB-231 cells stably expressing H2B-EGFP with PI staining