Fvb mice

FVB/NJ (8 wk old) female mice were used. Full thickness incisional wounds were generated in utero on embryonic day E15.5 as described previously [32 (link)] on three different dams. Briefly, a laparotomy was performed under aseptic conditions on time-mated FVB Mice (Taconic Biosciences; Hudson, NY, USA). Two-millimeter full thickness incision was made in the dorsal skin of fetus, using microsurgical scissors. One microliter of phosphate buffered saline containing 10% India ink (Becton Dickinson; Sparks, MD, USA) was injected subcutaneously at the wound site for subsequent wound identification. During the ultrasound imaging, the dorsal side of the fetus was scanned. The scattered or reflected ultrasound echoes were collected as an image signal. Based on the difference in acoustic impedance of different tissues the tissue types were identified. Wounds were with low echogenicity than normal tissue in ultrasound B-mode images [6 (link)]. The fetuses were euthanized for wound tissue collection at 48h post-wounding. All animal experiments were carried out with approval from Institutional Animal Care and Use Committee, The Ohio State University.

Male WT FVB mice (20–30 g, 8 weeks old), and mdr1a/1b^−/− (mdr1a/1b KO) and bcrp^−/− (bcrp KO) mice of the same genetic background (FVB) mice (20–30 g, 8 weeks old) were obtained from Taconic Farms (Germantown, NY, USA). Animals were maintained under standard conditions (temperature at 23 ± 1 °C with a relative humidity between 40% and 60%) with a 12 h light/dark cycle. Food and water were available ad libitum. All animal experiments were carried out in accordance with principles and procedures outlined in the National Institutes of Health Guide for the Care and Use of Laboratory. All experimental animal procedures were reviewed and approved by the Animal Care and Use Committee of the Kyoto Pharmaceutical University (2005-239) and Ritsumeikan University (BKC2010-27).

Experiments were conducted on 5 week old female FVB mice (Taconic Farms, Oxnard, CA). Mice were housed and treated in accordance with protocols approved by the University of Arizona Institutional Animal Care and Use committee (IACUC). On day zero, mice were anesthetized with an intramuscular injection of Ketamine/Xylazine (50 mg/kg/10 mg/kg respectively; Western Medical Supply, Arcadia, CA). Mice were then placed in a holding device, with the head and neck region exposed, while the remainder of the body was shielded with >0.6 mm of lead. Mice were irradiated with a single 5 Gy dose of radiation (day 0 of time course). On day four, a sub-group of untreated and irradiated mice received a single tail vein injection of mAbEDAR1 (5 mg/kg in PBS, Edimer Pharmaceuticals, Cambridge, MA) similar to a previous study using this compound [16] (link). The rationale for systemic (iv) administration of mAbEDAR included previous work on the functionality and half-life of mAbEDAR by this route, the feasibility of one injection reaching all major and minor salivary glands, and the greatest translatability to the clinical setting. The remaining untreated and irradiated mice received a vehicle (PBS) injection. All experiments were approved by IACUC.