Fig. 4

Rutaecarpine induces metabolic reprogramming in TNBC cells. A. KEGG analysis showing significant enrichment in metabolism, genetic information processing and human disease. B. Heat map of carbon metabolism and oxidative phosphorylation. C. ECAR and OCR test with TECAN for 1.5 h for the cells in 3D spheroids and quantification. Unpaired Student’s t-test; * p < 0.05; **** p < 0.0001. D. FCM test of SoNar-expressing cells in spheroids. Quantification of intracellular F488/F405 (NAD.⁺/NADH) ratios in single cells resuspended in HBSS buffer. Unpaired Student’s t-test; * p < 0.05. E. FCM analysis of PercevalHR-expressing cells in spheroids. Quantification of intracellular F488/F405 (ATP/ADP) ratios in single cells resuspended in HBSS buffer. Unpaired Student’s t-test; *** p < 0.001. F. Heat map of metabolomic analysis of DMSO- and rutaecarpine-treated MDA-MB-231 cells in spheroids. G. SCENITH test of the MDA-MB-231 cells in spheroids DMSO or rutaecarpine treatment. Student’s t-test; **** p < 0.0001