Fig. 7: Hesperidin is identified as a FADS inhibitor.

a Workflow for high-throughput virtual screening of FADS-targeting compounds. b Five compounds selected for experimental validation. c Proliferation of Hep3B, HCCLM3 and Hepa1-6 cells treated with the five compounds (100 µM) respectively, as determined by CCK-8 assay (n = 5 biological replicates). d Colony formation of Hep3B, HCCLM3 and Hepa1-6 cells following Hesperidin (100 µM) treatment. e, f Subcutaneous tumors generated by Hepa1-6 cells in Rag1^-/- mice (e) and C57BL/6 mice (f) (n = 5 per group). Hesperidin (100 mg/kg) or DMSO control was administered daily by intraperitoneal injection. Tumor volumes were measured at the indicated timepoints. Tumors were photographed and weighed at day 21. g Docking analysis showing the interaction between Hesperidin and FADS. h FAD levels in Hep3B and HCCLM3 cells treated with or without Hesperidin (100 µM, 48 h) (n = 3 biological replicates). i, j ROS and lipid peroxidation in Hep3B cells treated as indicated (n = 3 biological replicates). Hesperidin, 100 μM for 48 h; FAD, 20 μM for 48 h. k Percentage of dead cells (7-AAD⁺) in Hep3B cells (n = 4 biological replicates). Hesperidin, 100 µM for 48 h; FAD 20 µM for 48 h. l Representative images and quantification of CD8⁺ T cell density in subcutaneous Hepa1-6 tumors treated with or without Hesperidin (n = 5 per group). Data are presented as mean ± SD (c, e, f, i, j, k, l) or median with interquartile ranges (h). P-values were calculated using unpaired two-sided Student’s t test (e, f, h, i, j, k, l), and two-way ANOVA with Tukey’s test (c, e, f). Source data are provided as a Source Data file.