ATF4 overexpression induces weight gain and increases fat mass. (A) ATs were fed either a LFD or HFD for the indicated time periods. Serial body weight of each zebrafish was monitored monthly (n = 20–25). *p < 0.01, and **p < 0.005. (B) Representative images of male and female ATs − Dox and WT-Dox fed either a HFD for 5 months. (C) ATF4 overexpression leads to greater intra-abdominal fat (left), and the increased intra-abdominal fat pad size is shown (right). (D) Nuclear magnetic resonance intra-abdominal fat analyses of 10-month-old HFD–fed WT-Dox and AT1-Dox. Examined fish was prepared from 5–8 four months male–female fish pairs of each indicated groups fed either a HFD for 6 months. The asterisk represents statistically significant differences; *p < 0.01, and **p < 0.005. (E) Expression of adipogenic genes and proteins. PPAR-γ, C/EBP-β, and SREBP-1c mRNA (left, qRT-PC), and proteins (right, western blot). The qRT-PCRs were performed in triplicate in (E). Expression analysis of the selected genes using cDNA prepared from average 2–3 eight months male–female fish pairs per groups. 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.
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