Figure 1. Homozygous tp53^del/del zebrafish spontaneously develop a wide range of tumor types.

(A) tp53 genomic locus and CG1 tp53^del/del allele. TALEN arms were designed to target the 5’ and 3’ genomic sequence of tp53 (red). (B–M) CG1 tp53^del/del zebrafish develop leukemia (B–D), angiosarcoma (E–G), MPNSTs (H–J), and germ cell tumors (K–M). Whole animal images (B,E,H,K), hematoxylin/eosin (H and E) stained sections (C,D,F,G,I,L,M), and immunohistochemistry for Sox10 (J). Blast-like leukemia cells predominate in the kidney marrow and efface the renal tubules (black arrow, (D). (N) Tumor incidence in CG1 tp53^del/del zebrafish (n = 134). (O) Quantitation of tumor types that form in CG1 tp53^del/del mutant zebrafish by 55 weeks of life based on histology review (n = 51). (P–S) kRAS^G12D-induced embryonal rhabdomyosarcoma (ERMS) generated in CG1 tp53^del/del zebrafish. Whole animal bright field and GFP-epifluorescence overlap images (P and Q, respectively). H and E stained sections revealed features consistent with human ERMS (R,S). Scale bars equal 12.5 mm in whole animal images and 100 μm in histology images.

Figure 1—figure supplement 1. tp53^del/del zebrafish survive at expected Mendelian ratios, lack Tp53 protein expression, and are resistant to irradiation-induced cell death.

(A) Survival of animals by genotype from a heterozygous tp53^wt/del in-cross. tp53 homozygous wild-type (wt/wt), heterozygous (wt/del), and homozygous (del/del) mutant fish. (B) Western blot analysis at 24 hr post-fertilization (hpf) whole embryos. Actin is used as a loading control. (C–D) TUNEL staining performed on tp53^wt/wt and tp53^del/del embryos following gamma-irradiation at 24hpf (16 Gray) and fixation at 30hpf. Whole embryos images are shown for representative animals of each genotype (C). Quantification of TUNEL-positive cells within 1000 micron² area. Regions where cells were counted are outlined by the white boxes in panel C. p<0.001 by Student’s T-test.

Figure 1—figure supplement 2. Tumor latency in tp53^del/del zebrafish.

Designation as assessed by histology review.