A graphical timeline of conditioning is shown in Figure 1B . Mice were given 500μg Diphenhydramine HCl i.p. approximately 10–15 min prior to αCD117. 500μg αCD117 was injected retro-orbitally into mice under isoflurane anesthesia on day −6 prior to HCT. Mice were irradiated on day −3 with 200 cGy (Figures 1 and 2 ) or 300 cGy (Figures 3 and 4 ) TBI. 300μg each of αCD4 and αCD8 was administered i.p. on days −2, −1, and 0. αCD117 mAb, ACK2 was purchased from Bio X Cell (Lebanon, NH) or BioLegend (San Diego, CA). αCD4 (GK1.5) and αCD8 (YTS169.4) were purchased from Bio X Cell. Diphenhydramine HCl was purchased from MedChem Express (Monmouth Junction, NJ).
Animal irradiation (XRT) was performed using a Kimtron Polaris IC-250 Biological Irradiator (Oxford, CT) with a 225 kV X-ray tube filtered by 0.5mm Cu source set at 225kV, 13.3mA. Mice were divided in irradiation pie cages from Braintree Scientific (Braintree, MA) when irradiated. Dosimetry calibration for our setup was performed using published methods on radiochromic film (Ma et al., 2001 (link)).
Animal irradiation (XRT) was performed using a Kimtron Polaris IC-250 Biological Irradiator (Oxford, CT) with a 225 kV X-ray tube filtered by 0.5mm Cu source set at 225kV, 13.3mA. Mice were divided in irradiation pie cages from Braintree Scientific (Braintree, MA) when irradiated. Dosimetry calibration for our setup was performed using published methods on radiochromic film (Ma et al., 2001 (link)).