Figure 4—figure supplement 2.

Assigning pathogenicity to two human TP53 sarcoma mutations in the kRASG12D-driven embryonal rhabdomyosarcoma (ERMS) model.

(A) Tumor numbers per zebrafish expressing no TP53 (tp53^-/-), or TP53^C176F, or TP53^P153Δ.n = 36 (tp53^-/-), n = 58 (TP53^C176F), n = 81 (TP53^P153Δ). (B) Percentage of EdU-positive cells in representative areas of fish tumors expressing either no TP53 (tp53^-/-), TP53^C176F, or TP53^P153Δ. n = 8–10. (C) Total number of pHH3-positive cells per single ERMS tumor ×200 confocal image section assessed from tumors expressing WT TP53, TP53^C176F, or TP53^P153Δ intp53^-/- zebrafish; n = 3–5 primary tumors. (D) Representative images of tumor-burdened adult zebrafish (4–6 months of age) with tumors expressing kRAS^G12D + TP53^C176F (DsRed+) treated with either DMSO or ZMC1 over a span of 3 wk. (E) Western blot showing p53 expression levels in fish tumors expressing kRAS^G12D + TP53^C176F and treated with either DMSO or ZMC1. (F) Percentage of Annexin V-positive cells in tumors expressing kRAS^G12D + TP53^C176F that were treated with either DMSO or ZMC1. n = 4. (G) Ratio of tumor size to total body for in fish tumors expressing kRAS^G12D + TP53^C176F, treated with either DMSO or ZMC1. ns, not significant, n = 4. (H) Representative images of tumor-burdened zebrafish expressing kRAS^G12D (DsRed+) treated with either DMSO or ZMC1 over a span of 3 wk. Scale bar = 2 mm (I) Western blot showing p53 expression levels in Rh30 cells (control) and fish tumors expressing kRAS^G12D, treated with either DMSO or ZMC1. (J) Percentage of Annexin V staining of fish tumors expressing kRAS^G12D, treated with either DMSO or ZMC1. ns, not significant. n = 6–7. (K) Ratio of tumor size to total body in tumors expressing kRAS^G12D, treated with either DMSO or ZMC1. ns, not significant, n = 4.