Quantitative RT-PCR and western blot analysis.
Samples were prepared from (A, C) 4-dpf whole embryos, and (B, D) internal organs from 3-month-old zebrafish. Data are means, and bars indicate standard error. (A, B) Quantitative RT-PCR showed higher upregulation of inflammation-related genes in Sirt1-/- than in controls. (C, D) Western blot hybridization showed upregulation of IL-1b, active-caspase 3 and TGF-β. Control, wildtype; IL-1b, 18kDa; caspase 3a, 29 and 31 kDa; TGF-β, 45 kDa; β–actin, 43 kDa. *P < 0.05 versus control.