Chromatogram and UV and MS spectrum data of phenolics in miracle berry leaf extract. 1. gallic acid; 2. 5-caffeoylquinic acid; 3. 5-O-p-coumaroylquinic acid; 4. 3-O-p-coumaroylquinic acid; 5. 3-O-feruloylquinic acid; 6. 4-O-caffeoylquinic acid; 7. 4-O-p-coumaroylquinic acid dimer; 8. 4-O-p-coumaroylquinic acid; 9. 4-O-feruloylquinic acid; 10. myricetin-3-galactoside; 11. rutin; 12. myricetin-3-O-rhamnoside; 13. quercetin-3-D-galactoside; 14. quercetin-3-glucoside; 15. kaempferol-3-O-glucoside; 16. quercetin-3-rhamnoside; 17. myricetin-O-galloyl rhamnoside; 18. quercetin 3-O-α-(2”-galloyl) rhamnoside.

Mortalities of zebrafish embryo at different concentrations of MBL extract (A), inhibition rates of subintestinal vein (SIV) developments (B), and fluorescent images of zebrafish embryos (C) at different MBL extract concentrations. Bars with different letters indicate a significant difference (p < 0.05).

Effect of PTX and MBL extract on the growth of MCF-7 tumors. (A) Growth curves of MCF-7 xenograft tumors from different treated groups after treatment. (B) Photographs of all xenograft tumors in mice.

Apoptosis of tumor cells induced by PTX and MBL extract was detected by flow cytometry. (A) Tumor cells were stained with Annexin V-FITC/PI for flow cytometry analysis. Apoptotic cells were divided into 2 stages (LR for early apoptotic cells and UR for late apoptotic cells). (B) The total apoptotic rates examined by flow cytometry, including the early and late apoptosis. *p < 0.05.

Metabolomics analysis of MCF-7 xenograft mice serum. (A) PCA scores of serum metabolites from NC, NT, PTX, LSD, and HSD group. (B) OPLS-DA scores plots of MCF-7 xenograft mice and normal control. (C) Hierarchical clustering of serum metabolome. The heat map represented the Z scores of significantly differentially expressed metabolites between the NC and the NT group. (D) ROC curves for the combination of serum succinic acid and glucose-6-phosphate to discriminate MCF-7 xenograft mice from normal control. (E) Enriched KEGG pathway in 42 up-regulated (red) and 37 down-regulated (green) metabolites of NT group compared to the NC group. The x axis shows the enrichment significance presented with –log2 (P-value).

Metabolomics analysis of PTX, LSD, and HSD treated MCF-7 xenograft mice. (A) Heatmap shows the commonly changed metabolites in the treatment groups compared to the NT group. The data were represented as the log2 of fold change (treatment/NT). (B) The enriched metabolic pathways in PTX, LSD and HSD groups compared to NT group. The heat map represented –log2 (P-value) of enrichment significance. White square means pathway term was not enriched.

Co-regulated metabolic pathways in PTX and MBL extract treated MCF-7 xenograft mice. (A) Altered unsaturated fatty acids metabolic pathway. (B) Altered purine metabolism. Boxes represented for the level of metabolites identified in NC (green), NT (blue), PTX (red), LSD (purple), and HSD (orange) group. Data represent means ± standard deviation. * is represent statistical significance with P < 0.05 when compared to NT group.