Figure 1

Figure 1

Establishing Tg (cyclin B2:Luc/GFP) MITO-Luc/GFP transgenic zebrafish lines. (A) Schematic representation of the pT2KXIGΔin-cyclin B2-Luc/GFP expression vector. A fragment of murine cyclin B2 promoter containing CCAAT boxes (black) was cloned upstream of the firefly luciferase gene (vertical stripes) and the green fluorescent protein GFP (horizontal stripes), in the pT2KXIGΔin plasmid containing the Tol 2 transposable element (gray). The binding of NF-Y to the CCAT boxes is shown. (B) Relative luciferase activity obtained in one to four-cell stage AB embryos transiently injected with the pT2KXIGΔin-Luc/GFP or the pT2KXIGΔin-cyclin B2-Luc/GFP. The error bars are mean standard deviations from two experiments performed in duplicate. Asterisks denote significant differences between pT2KXIGΔin-cyclin B2-Luc/GFP and empty vector assessed by t-test (*p < 0.05). (C) Quantification of transgene copy number in MITO-Luc/GFP zebrafish lines by quantitative PCR. Amplification of 10/15 transgene copies from MITO-Luc/GFP¹ and 1 from MITO-Luc/GFP² genomic DNA, is quantified by comparing amplifications from known pT2KXIGΔin-cyclin B2-Luc/GFP copy numbers. Full-length gel is presented in Supplementary Figure S17. (D) Relative luciferase activity in MITO-Luc/GFP¹ and MITO-Luc/GFP² zebrafish lines. The error bars are mean standard deviations from two experiments performed at least in triplicate with 3 embryos in each group. Asterisks denote significant differences between MITO-Luc/GFP¹ and MITO-Luc/GFP² embryos assessed by t-test (****p < 0.0001).