(A to F) Western blots (A) of cardiac lysate from transgenic mice show increased DDiT4L abundance (B), decreased phosphorylation (p) of Thr389 in S6K (C), increased LC3II/I ratio (D), decreased phosphorylation of Ser757 in Ulk1 (E), and no changes in p62 abundance (F). These effects were reversed upon transgene suppression. n=6–9 mice per genotype. One-Way ANOVA, Bonferroni posttest *p<0.05, **p<0.01, ****p<0.0001. Representative Western Blots are from the same blot, but they may not be contiguous Phospho-proteins were normalized to total protein.
(G to I) Mice overexpressing DDiT4L from birth show decreased fractional shortening (FS) (G), thinner ventricular walls (H) and a larger left ventricular diastolic dimension (LVDd) (I) which was reversed upon doxycycline administration. n=6–9 mice per genotype. Two-Way ANOVA, Bonferroni posttest. *p<0.05 compared to Tta control, #p<0.05 compared to DDiT4L/Tta.
(J) ANP expression was increased and the A/B MHC ratio and Cited4 expression was decreased in mice overexpressing DDiT4L which was reversed with transgene suppression. n=6–9 mice per genotype. One-Way ANOVA, Bonferroni posttest, *p<0.05 compared to control or indicated sample.
(K) EM images showed increased numbers of autophagosomes (indicated by the arrows) in cardiomyocytes from DDiT4L overexpressing mice. n=4 mice per genotype. Scale bar represents 5 μM.