FIGURE SUMMARY
Title

Identification of benzodiazepine Ro5-3335 as an inhibitor of CBF leukemia through quantitative high throughput screen against RUNX1-CBFbeta interaction

Authors
Cunningham, L., Finckbeiner, S., Hyde, R.K., Southall, N., Marugan, J., Yedavalli, V.R., Dehdashti, S.J., Reinhold, W.C., Alemu, L., Zhao, L., Yeh, J.R., Sood, R., Pommier, Y., Austin, C.P., Jeang, K.T., Zheng, W., and Liu, P.
Source
Full text @ Proc. Natl. Acad. Sci. USA

NSC140873, Ro5-3335, and Ro24-7429 inhibit definitive hematopoiesis in zebrafish embryos. (A) NSC140873 reduced circulating cd41-GFP+ (Upper) and lck-GFP+ (Lower) cells in transgenic zebrafish embryos compared with DMSO-treated embryos. Red arrows point to circulating cd41-GFP+ cells in the heart and lck-GFP+ cells in the thymus, respectively. (B) Dose-dependent reduction of circulating cd41-GFP cells by NSC140873. (C) Chemical structures of the three related compounds. (D) Dose-dependent reduction of circulating cd41-GFP cells by Ro5-3335 and Ro24-7429. (E) The runx1 W84× truncation mutation sensitizes fish embryos to the inhibition by Ro5-3335. *Numbers of untreated runx1+/- embryos or DMSO-treated runx1+/- embryos with intermediate or no visible circulating CD41-EGFP+ cells were compared. #Numbers of runx1+/- embryos or wild-type embryos with intermediate or no visible circulating CD41-EGFP+ cells after Ro5-3335 treatments were compared. For all comparisons in B, D, and E, the P values were calculated with 2 × 2 contingency tables and Fisher’s exact test.

Ro5-3335 rescues hematopoietic defect in the RUNX1ETO transgenic embryos. (A) RUNX1-ETO transgenic embryos at 48 h postfertilization (hpf). HS, heat shock. Arrows point to the posterior blood island. Note the pooling of blood cells in the heat-shocked embryo. (B) Heat-shocked RUNX1-ETO transgenic embryos at 48 hpf that were treated with either 25 μM Ro5-3335 (Upper) or DMSO (Lower). (C) Bar graphs showing the effect of incubating heat-shocked RUNX1-ETO transgenic embryos with Ro5-3335 at the indicated concentrations. Black bars, embryos with blood pooling and no circulation; gray bars, embryos without blood pooling but also no circulation; white bars, embryos without blood pooling and with circulating cells. Means of three independent experiments are shown with SDs. *P < 0.03; **P < 0.003.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Proc. Natl. Acad. Sci. USA