FIGURE SUMMARY
Title

Loss of Ewing sarcoma EWS allele promotes tumorigenesis by inducing chromosomal instability in zebrafish

Authors
Park, H., Galbraith, R., Turner, T., Mehojah, J., Azuma, M.
Source
Full text @ Sci. Rep.

Loss of ewsa promotes tumorigenesis.

Total numbers of fish for each genotypes are: ewsaw/w;tp53w/w (n = 41), ewsaw/m;tp53w/w (n = 135), ewsam/m;tp53w/w (n = 21), ewsaw/w;tp53w/m (n = 28), ewsaw/m;tp53w/m (n = 46), ewsam/m;tp53w/m (n = 39), ewsaw/w;tp53m/m (n = 48), ewsaw/m;tp53m/m (n = 12), ewsam/m;tp53m/m (n = 19). (A) Representative images of zebrafish. In images of ewsa and tp53mutants, tumors are indicated with an arrow (>). Scale bar: 1cm. (B) Tumor incidence is higher in ewsaw/m;tp53w/m and ewsam/m;tp53w/m compared to ewsaw/w;tp53w/m; and in ewsaw/m;tp53m/m and ewsam/m;tp53m/m compared to ewsaw/w;tp53m/m zebrafish. (C) Average age at tumor onset is lower in ewsaw/m;tp53w/m and ewsam/m;tp53tp53w/m compared to ewsaw/w;tp53w/m; and in ewsaw/m;tp53m/m and ewsam/m;tp53m/m compared to ewsaw/w;tp53m/m zebrafish. (D) Representative images of H&E staining of tumors observed in zebrafish. MPNST-like: malignant peripheral nerve sheath tumor-like; SRBCT: small round blue cell tumor.

Loss of ewsa promotes chromosomal instability (CIN).

(A) Representative images of mitotic cells from 27 hours post fertilization (hpf) zebrafish embryos visualized by immunohistochemistry using an anti-α-tubulin antibody (red) and DAPI (blue). (B) Percentages of cells with aberrant mitosis in zebrafish embryos at 27 hours post fertilization, as determined after immunohistochemistry using an anti-α-tubulin antibody. Experiments were repeated three times. (n = 13 to 60 mitosis from 3 to 6 embryos per genotype). *P < 0.05. (C) Representative images of chromosomes obtained from metaphase spread of 24 hpf ewsaw/w (top) and MZ ewsam/m (bottom) embryos. (D) Numbers of chromosomes per cell, and percentage of cells from ewsaw/w (blue bars) and MZ ewsam/m (red bars) zebrafish embryos containing that number of chromosomes. The vast majority of ewsaw/w cells contained the normal chromosome number (50) (blue), whereas chromosome numbers in MZ ewsam/m mutant zebrafish cells varied from 34 to 56 chromosomes per cell (red). P < 0.05. Note that all samples were obtained from lines with wildtype tp53w/w background.

Acknowledgments
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