Pacgfp vector

Mouse LYVE‐1 cDNA was reverse transcribed with a First Strand cDNA Synthesis kit (GE Healthcare, Chicago, IL, USA) from total RNA prepared by Isogen II (Nippon Gene, Toyama, Japan) of mouse stomach tissues rich in lymphatic vessels. Green fluorescent protein (GFP) was fused to the carboxy terminus of full‐length (membrane form) or the extracellular domain (secretory form) of mouse LYVE‐1 in the pAcGFP vector (BD Biosciences, Mountain View, CA, USA). Transfection of the full‐length or secretory mouse LYVE‐1‐GFP vector into cells was performed using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. Cells were selected using culture media containing 400 μg/mL G418 (Nacalai Tesque, Kyoto, Japan), and were sorted by cellular green fluorescence using a JSAN cell sorter (Bay Bioscience, Kobe, Japan). These experiments were approved by the Safety Committee for Recombinant DNA Experiments at Kindai University (KDPS‐19‐002, KDPS‐21‐001 and KDPS‐26‐004).

GFP was fused to full‐length macaca LAT1 in a pAcGFP vector (BD Biosciences, Mountain View, CA, USA). Transfection of macaca LAT1‐GFP vector into RH7777 or HEK293 cells was carried out using Lipofectamine 3000 (Invitrogen). Cells were selected with 400 μg/mL G418 (Nacalai Tesque, Kyoto, Japan), and clone‐sorted for cellular green fluorescence using a JSAN cell sorter (Bay Bioscience, Kobe, Japan).