Cremaphor el

SBFI-102, SBFI-103, and docetaxel were each reconstituted in a 1:1:8 vehicle consisting of dimethyl sulfoxide (DMSO) (Thermo Fisher Scientific, Hampton, NH):Cremaphor-EL (Sigma-Aldrich):saline. SBFI-102 and SBFI-103 were administered via intraperitoneal injection (ip) using a 27G needle at 20 mg/kg daily. Docetaxel was administered i.p. at 5 or 10 mg/kg weekly. All drugs were administered in a volume of 10 μL/g body weight.

All mice received 3 intraperitoneal (i.p.) injections of bromodeoxyuridine (BrdU; 150mg/kg dissolved in 0.9% saline and 0.007 N NaOH; Boehringer Mannheim), one injection every 6 hours to label dividing cells in S-phase. 2 days following BrdU injections, mice were administered P7C3 (provided by Andrew Pieper, University of Iowa, Iowa City, IA) or vehicle as follows: P7C3 was dissolved in 2.5% dimethyl sulfoxide (DMSO) transferred to a glass vial containing 10% Cremaphor EL (Sigma, C5135, St. Louis, MO) and vortexed vigorously for 1 minute. Sterile-filtered 5% dextrose (pH 7.0) was added to create a 1.25mg/mL working solution of P7C3 and vortexted vigorously. P7C3 was prepared fresh daily and stored at 4°C until use. Vehicle (Veh) consisted of 5% dextrose (pH 7.0) and 10% Cremaphor EL. Mice were administered 350μL (437.5μg) of P7C3 or Veh twice daily via i.p. injections between 8–9am and 5:30–7pm for 90 days.

(+)—WIN55,212-2 and (−)—CP55,940 were purchased from Cayman Chemical (Ann Arbor, MI). CP55,940 was prepared in 100% ethanol and subsequently diluted 20-fold in vehicle containing 0.9% saline and 5% Cremaphor EL (Sigma-Aldrich, St Louis, MO) (18:1 vehicle v/v). WIN55,212-2 was prepared in 100% DMSO and diluted in vehicle containing 0.9% saline, 5% Cremaphor EL, and 5% ethanol (18:1:1 vehicle v/v/v) for injection. The total amount of DMSO contained in all WIN55,212-2 injections was 4% v/v. Cisplatin was purchased from Tocris (Minneapolis, MN) and subsequently dissolved in 0.9% saline. Sodium bicarbonate (Fisher Scientific, Pittsburgh, PA) was dissolved in 0.9% saline to prepare a 4% w/v solution for administration. Drugs were administered via intraperitoneal (i.p.) injection (CP55,940, WIN55,212-2, Cisplatin) in a single volume of 10ml/kg of body weight or via subcutaneous (s.c.) injection (Sodium bicarbonate) in a single volume of 1 ml. Formalin (2.5% v/v) was prepared by diluting formaldehyde (37%, Fisher Scientific) in water.