Hinokitiol

Male 8-10-week-old BALB/c mice were obtained from Nihon CLEA Tokyo, Japan. Mice were maintained under standard conditions in accordance with our institutional guidelines. All animal experiments were approved by the Institutional Animal Care and Use Committee of Niigata University (SA00223). Hinokitiol was purchased from FUJIFILM Wako Pure Chemical Industries (Osaka, Japan) and solubilized in phosphate- buffered saline (PBS) containing 10% ethanol. Thiazolyl blue tetrazolium bromide (MTT) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The anesthetic mixture consisted of medetomidine hydrochloride (Domitol; Meiji Seika Pharma Co., Ltd., Tokyo, Japan), midazolam (Dormicum; Astellas Pharma Inc., Tokyo, Japan), and butorphanol (Vetorphale; Meiji Seika Pharma Co., Ltd.) mixed at 0.15, 2.0, and 2.5 mg/kg body weight with normal saline (Otsuka Pharmaceutical Factory, Inc. Tokyo, Japan) [22 ].

Human holo‐Tf (208‐18971) and hinokitiol (085‐06251) were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Recombinant human EPO (873999) was purchased from Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan). Percoll (17‐0891‐02) was obtained from GE Healthcare (Buckinghamshire, UK). FITC‐anti‐mouse CD44 (103006), PE‐conjugated anti‐mouse Ter119 (116208), PE/Cy7‐conjugated anti‐mouse CD71 (113812), Pacific blue‐conjugated anti‐mouse CD45 (103126), biotin‐conjugated anti‐mouse CD45 antibodies (103104), 7‐aminoactinomycin D (7‐AAD; 420404), MojoSort™ Streptavidin Nanobeads (480016), and MojoSort™ Magnet (480019) were purchased from BioLegend (San Diego, CA, USA). The anti‐TfR1 monoclonal antibody R17 208.2 (sc‐65883) was obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Control rat IgM (14‐4341‐85), SYTO16 (S7578), and Cell‐Tak Cell and Tissue Adhesive (354240) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). RITC‐conjugated anti‐rabbit IgG antibody (SA00007‐2) was obtained from Proteintech (Rosemont, IL, USA). MitMAB (ab120466) and anti‐TfR1 antibody (ab84036) were obtained from Abcam (Cambridge, UK).