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. 2018 Jun 5;7:e33800. doi: 10.7554/eLife.33800

Figure 1. Zebrafish models of human PAX3-FOXO1 tumorigenesis.

(A) Zebrafish were injected at the single-cell stage with mosaic GFP2A-tagged human PAX3-FOXO1 under the control of various promoters. At 24 hr old, embryos were sorted for GFP expression indicating successful injections (typically 99% GFP+) and were allowed to grow and monitored for up to 19 months to develop tumors. (B) Beta-actin-driven PAX3-FOXO1 primarily produced primitive neuroectodermal tumors in a wild-type genetic background. Shown for all tumors are representative examples with the presentation of gross morphology and GFP expression patterns coupled with a hematoxylin and eosin stain. (C) CMV-driven PAX3-FOXO1 produced rhabdomyosarcoma in the tp53M214K/M214K-sensitized genetic background. (D) Ubiquitin-driven PAX3-FOXO1 produced an undifferentiated sarcoma in a wild-type genetic background. (E) RNAseq data from zebrafish PAX3-FOXO1 fluorescent tumors showing the number of reads supporting the presence of the human fusion-oncogene. (F) Tumor incidence of GFP + tumors detected in BetaActin-GFP2A-PAX3FOXO1 (n = 74) injected zebrafish versus BetaActin-GFP (n = 147) injected controls in a wildtype genetic background. (G) Tumor incidence of GFP + tumors detected in CMV-GFP2A-PAX3FOXO1 (n = 31) injected zebrafish in a tp53M214K-sensitizing genetic background.

Figure 1.

Figure 1—figure supplement 1. Promoter restricted expression of human PAX3-FOXO1 has different effects on survival in developing zebrafish.

Figure 1—figure supplement 1.

Shown are the survival curves for the following promoters and the total number of zebrafish embryos analyzed for each condition: beta actin (n = 120), mitfa (n = 45), fli1 (n = 81), ubi (n = 160), unc503 (n = 108), cmv (n = 20), SA-splice acceptor (n = 153).
Figure 1—figure supplement 2. Junctional sequence used to map human PAX3-FOXO1 fusion RNAseq reads from zebrafish tumors.

Figure 1—figure supplement 2.