Urethane ethyl carbamate

All salts for Krebs solutions were of AnalaR grade or higher and obtained from BDH, Poole, or Fisons, Loughborough. Chloroquine diphosphate, (±) adrenaline HCl, urethane (ethyl carbamate), Tween 80, N,N‐dimethylacetamide and propylene glycol were obtained from Sigma, Poole. Heparin sodium (Multiparin of 5,000 IU·ml⁻¹) injection was obtained from CP Pharmaceuticals, Wrexham. MTBE was purchased from Fisons, Loughborough; Ro5‐4864 was purchased from Fluka Chemika, Gillingham, Dorset; Hypnorm (fentanyl/fluanisone) was from Janssen Animal Health, Petteridge, Kent; Amprol Plus (Amprolium HCl/ethopabate) was from Merk Sharp & Dohme, Hertfordshire; and sodium pentobarbitone was from RMB Animal Health Ltd, Dagenham. 7‐Chloro‐4‐(5‐diethylamino‐1‐methylpentyl‐amino)‐quinoline diphosphate was a gift from the Walter Reed Army Institute of Research, Washington D.C.; and clonazepam and diazepam were gifts from Hoffman‐La Roche, Basel.

All animal care and experimental procedures were approved and conducted according to guidelines issued by the Vanderbilt University Institutional Animal Care and Use Committee. MerTK-KO mice on FVB background were provided by Dr. R. Cook (Vanderbilt University, Nashville, TN) [40 (link)]. Lung tumors were induced in IKKβ^Δmye mice (IKKβ^fl/fl; LysM-Cre) [41 (link)] and littermate WT controls by a single intraperitoneal (i.p.) injection of urethane (ethyl carbamate, 1 g/kg) (Sigma-Aldrich). BAY 11-7082 (10 mg/kg body weight; Cayman Chemical) was delivered by i.p. injection as described previously [27 (link)]. Warfarin (250mg/L, Sigma) was delivered with drinking water. Lung tumors were induced in LSL-Kras^G12D mice [42 (link)], using intratracheal (i.t.) instillation of adeno-Cre (1.5 × 10⁷ plaque-forming units). Subcutaneous tumors in C57BL/6 mice were established by an injection of 5 × 10⁵ syngeneic LLC cells in 100 ul of PBS in the right flank. When tumor size reached about 3 mm in diameter (day 6), mice were randomized and treated with vehicle control, BAY 11-7082 (10 mg/kg, i.p.,), Foretinib (30 mg/kg, gavage), or a combination of BAY 11-7082 and Foretinib every second day. Tumor sizes were measured using Traceable digital calipers (Fisher Scientific). Bronchoalveolar lavage (BAL) cells were collected and counted as previously described [12 (link)].