Fig 1. Dynamic autophagy signaling in a zebrafish adult AIC (aAIC) model and modifying effects of atg7+/−.

(A) Schematics of the experimental procedure of an anthracycline-induced cardiotoxicity (AIC) in an adult (aAIC) zebrafish model (DOX, doxorubicin). (B) Dynamics of EF% in the DOX-treated zebrafish and the control group using a high-frequency echo system (n=15). (C) Representative Western blot showing temporal changes in LC3-II protein expression in the hearts of adult zebrafish with AIC. Bafilomycin A1 (30 nM) was administered 4 h before the zebrafish were sacrificed. (D to G) Quantification of LC3-II and the ratio between hearts treated with and without BafA1 in (C), n=3 hearts/group. (H) Representative images of a Western blot showing the LC3 expression in the hearts of WT and atg7+/− zebrafish in the absence or presence of 30 nM BafA1 for 4 h. (I) and (J) Quantification of the Western blot data in (H), n = 3 in each group. (K) Ventricular ejection fraction of WT and atg7−/+ zebrafish with or without DOX stress 8 weeks post injection (wpi) (n=10 fish/group). (L) Quantification of nppa and nppb gene expression by quantitative RT-PCR. n=3 per group. Student’s t test was used in (B); Mann-Whitney test in (E), (G) and (J); Kruskal-Wallis test in (D), (F), (I), and (L); one-way ANOVA followed by post hoc Tukey’s test in (K).