Transit ee delivery solution

C57BL/6J mice were obtained from CLEA Japan, Inc. The experiments were performed according to the guideline set by the institutional animal care and use committee of the University of Tokyo. Hydrodynamic gene delivery was allowed the standard procedure of TransIT-EE Delivery Kit (Mirus Bio). Twenty micrograms of purified plasmid (pLIVE-YB-1 or pLIVE-LacZ control plasmid) was dissolved in 1.8 ml of TransIT-EE Delivery Solution (Mirus Bio) and then was injected via mouse tail vein.

We used 5–6-week-old Rosa26^Ai14/WT mouse or Rosa26^{PA-Cre B4/Ai14} mouse. The abdominal surface furs of mice were removed using hand trimer. The fur-removed mice were rested for at least 24 h in a cage, and intraperitoneally injected with cDNAs encoding CAG-Magnets, CAG-Magnets-opti, Cre, or pCAG-Flpe (Addgene, #13787) using TransIT®-EE Delivery Solution (Mirus Bio LLC), respectively, according to the manufacturer’s protocol. The amount of injected DNA was 10 μg per mouse. The volume of delivery solution was 0.1 mL per mouse weight (g). After the hydrodynamic injection of DNA, the mice were kept in the dark for 8 or 32 h and then illuminated with a LED source (470 ± 20 nm; 200 W/m², CCS) for 16 h in a cage. After illumination, the mice were kept in the dark to recovery for 24 h. As with ambient light illumination, the mice were kept under a standard white fluorescent light (0.05 W/m²) for 48 or 72 h. The mice were sacrificed using standard procedure approved in our animal protocol and their livers were immediately harvested for fluorescent imaging with fluorescent stereoscope (Zeiss).