Fig. 5.

Characterization of A11. (A) The A6, A8, A10, and A11 compounds are analogs (structures in Supplementary Fig. S2). They showed different degrees of heart failure attenuation in the AA model. A11 seems to be the most potent among them. (B) Dosage curve of A11. The prime attenuation of heart failure is observed at nearly 90% by 10 μM. (C–E) A11 can cause albinism throughout the pigmentation stages. Embryos treated with 10 μM A11 at the beginning (C, 24–55 hpf) or during (D, 36–55 hpf) or after (E, 48–55) pigmentation showed defects in melanocytes and different degrees of albinism. Unlike MEK-I, A11 can reduce the existing melanin and/or melanocytes. Another difference between A11 and MEK-I is that A11 does not cause any notochord defects. (F–H) A11-induced albinism is dosage dependent. (I–K) A11 analogs also show different degrees of albinism. The albinism and heart failure attenuation ability of the A11 analogs seems to show a positive correlation, as A11 with the strongest albinism and heart failure attenuation and A10 with the least in both. (L) Time course experiment shows that the critical time window of A11 function is between 24 (∼80% attenuation) and ∼40 (∼40% attenuation or 50% efficacy) hpf. Paired t-test comparing 24 hpf and other time points showed P=0.286 (30 hpf), 0.011 (36 hpf), 0.019 (42 hpf), and 0.007 (48 hpf). The data presented are means±SEM of triplicate wells (n=3), and the dosage curve experiments were repeated at least thrice.