Fig. 5

Fig. 5

ATF4 overexpression sensitizes zebrafish to oxidative stress induced by a HFD and induces NASH. (A) Levels of hepatic MDA, and (B) H₂O₂ in hepatic mitochondria was compared between AT1 ± Dox and WT ± Dox adults fed a HFD or LFD for 4 weeks. (C) Representative histological analysis of livers from the AT1 ± Dox and WT ± Dox adults that were fed a HFD or LFD for 4 weeks (400X magnification, scale bars: 10 μm). (D) Molecular analysis of AT1-Dox NASH-like livers revealed the upregulation of inflammatory genes, il-1b, il-6, tnf-α, ifn-γ, nfkb2, and NF-kB. (E) Molecular analysis of AT1-Dox NASH-like livers revealed the upregulation of ER stress markers, atf6, ern2, ire1, prek, hspa5, grp78, and ddit3. The biochemical analysis of oxidative stress were performed in triplicate in (A,B) and prepared from average 2–3 eight months male–female fish pairs per groups (WT-Dox, WT + Dox, AT1-Dox, AT1 + Dox). The qRT-PCRs were performed in triplicate in (D,E). Expression analysis of the selected genes using cDNA prepared from average 2–3 eight months male–female fish pairs per groups (WT-Dox, WT + Dox, AT1-Dox, AT1 + Dox). Levels of mRNA were normalized to β-actin and expressed as fold of values in the WT-Dox control. The asterisk represents statistically significant differences; *p < 0.01, and **p < 0.005.