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. 2019 Jun 4;8:e44927. doi: 10.7554/eLife.44927

Figure 1. Generation of OF mice.

Figure 1.

(A) Schematic representations of the hypocretin gene, targeting vector, and targeted gene. To achieve orexin neuron-specific expression of Flp recombinase, we inserted EGFP-2A-Flp just behind the translation initiation site of the hypocretin gene in-frame. Viral T2A peptide is cleaved just after translation, and EGFP and Flp recombinase localize independently. DT, diphtheria toxin; Neo, neomycin-resistant gene expression cassette. (B) Structure of the reporter gene in the presence of Flp. Orexin-Flp; FSF-mTFP1 bigenic mice were generated to express mTFP1 in orexin neurons. (C) Representative pictures from coronal brain sections of Orexin-Flp; FSF-mTFP1 bigenic mice. Arrow indicates mTFP1 and/or EGFP expressing orexin neurons and the arrowhead indicates the position of the MCH neuron. Orexin-IR, orexin-immunoreactive; MCH-IR, MCH-immunoreactive. Inset scale bar: 20 µm. (D) Summary of the co-expression analysis (n = 4 mice). The p values were determined using one-way ANOVA test followed by a post-hoc Tukey analysis. Data represent the mean ± SEM.

Figure 1—source data 1. Source data for Figure 1D.
elife-44927-fig1-data1.xlsx (11.6KB, xlsx)
DOI: 10.7554/eLife.44927.003