Extended Data Fig. 4: MAT2A inhibition by FIDAS-5 induces senescence in liver cancer cells.
a, Cell viabilities of Huh7 and Hep3B cells treated with indicated doses of FIDAS-5 for 72 hours. b, Huh7 and Hep3B cell growth curves in the presence of 1 µM FIDAS-5. c, Relative SAM and SAH abundance determined by LC-MS analysis of control (n = 4) and FIDAS-5 (n = 4) treated Huh7 cells for 72 hours. d, Cell death quantification (Annexin V+%) of Huh7 and Hep3B cells with or without 72-hour FIDAS-5 exposure (Huh7: 5 µM; Hep3B:1 µM). e, Representative immunofluorescence images of vehicle control and FIDAS-5-induced senescent (FIS) Huh7 cells stained with 53BP1 (red), γH2AX (red) and DAPI (blue). Scale bar: 50 µm. f, Quantification of cells with >2 53BP1 foci in control (n = 5 200x fields) and FIS (n = 5 200x fields) Huh7 cells. To quantify, 5 random fields containing at least 30 cells were counted and averaged. g, Representative crystal violet staining of control and FIS Huh7 cells replated in growth medium for 3-week clonogenicity assays. h, qRT-PCR analysis of indicated gene expression in control and FIS Huh7 cells. i, j, Representative SA-β-gal staining (i) and quantification (j) of vehicle control (Veh) and FIDAS-5 treated TOV21G cells. To quantify in (j), 5 random 100x fields were counted and averaged. n = 5 200x fields for each group. Scale bar: 100 µm. k, Mouse body weight curves over time in vehicle control (Veh, n = 7 female BAB/c mice) and FIDAS-5 (n = 5 female BAB/c mice) treatment groups. Data presented as mean ± s.e.m. of three independent experiments (a,b,d,f,h,j,k) or mean ± s.e.m.(c); statistical significance was determined by a two-tailed Student’s t-test (a-d,f,h,j,k). Experiments were repeated three times independently, with similar results (e,g,i).
