Cld 8909

Mice were maintained under specific pathogen‐free conditions and experimental protocols approved by the University of Cincinnati IACUC. For prostate cell injections, 1.0 × 10⁶ cells were injected subcutaneously into the flanks of 6−8‐week‐old male FVB mice as described.¹⁹, ²⁰ Tumor growth was measured via calipers and volume was determined by the formula 0.5 × Length × Width².²¹ Surgical castration was performed as described when tumors reached 1000 mm³.¹⁹, ²⁰ For in vivo kinase inhibitor studies, mice were treated with 50 mg/kg/day BMS‐777607 (Selleck Chemicals) or methylcellulose (vehicle) via oral gavage. Mice treated with clodrosome (Encapsula Nanosciences CLD8909) for macrophage depletion were injected with 200 µl intraperitoneally every 3 days. Hi‐Myc mice (FVB‐Tg(ARR2/Pbsn‐MYC)7Key/Nci, Strain # 01XK8) were obtained through the mouse repository at the National Cancer Institute and crossed with ARR₂Pb‐RON strain B mice which were described previously.¹², ²² Mice were euthanized and tissues were collected for analysis at 30 weeks of age.

Chimeric mice were produced by adoptive transfer of donor bone marrow cells into irradiated recipient animals using combinations of wild‐type (WT; C57BL/6NJ) and KO (IRG1^−/−) mice in the following recipient/donor combinations: WT/WT, WT/KO, KO/WT, KO/KO. Recipient mice (6‐8 weeks) were injected intraperitoneally with 200 μL of liposome‐encapsulated clodronate (CLD‐8909; Encapsula NanoSciences) 24 hours prior to irradiation. Recipient mice were then exposed to an otherwise lethal 1,000 cGy from a cesium source (Nordion International) 6 hours before receiving 5 × 10⁶ bone marrow cells by tail vein injection. The bone marrow cells were prepared in a sterile manner from the tibia and femur bones of the donor mice. All animals were monitored 3 times weekly for the first 2 weeks to ensure successful bone marrow engraftment. The chimeric mice underwent hepatic I/R after 8‐12 weeks to ensure stable engraftment.