Clozapine

Troglitazone, clozapine, tienilic acid, and acetaminophen were purchased from Tocris Biosciences, Enzo Life Sciences, Sigma-Aldrich, and Sigma-Aldrich, respectively, and used without further purification. Other compounds and probes were synthesized and routes and analytical characterization are described in Supplementary Material. Mouse liver S9 fraction was obtained as an equal mix of male and female, samples from Xenotech, Lenexa, KS. All mouse studies were performed following protocols that received approval from The Scripps Research Institute–Institutional Animal Care and Use Committee office.
See Supplementary Methods sections for detailed experimental procedures.

All experiments using laboratory animals were in compliance with the Guidelines of the Canadian Council on Animal Care, and previously approved by the Animal Care Committee at the University of British Columbia. Adult male Sprague-Dawley rats were purchased at Charles-River (Montreal, QC, Canada), and housed under standard conditions of temperature (22±1ºC), humidity (70%), and light/dark (12/12-h cycle), with unlimited access to standard rat chow pellets and drinking water. Haloperidol hydrochloride (1 mg/kg/day) and clozapine (20 mg/kg/day) were both from Tocris (Bristol, UK), and dissolved in 0.9% NaCl saline solution adjusted to pH 5.5. Animals (n = 10 per group) received one daily intraperitoneal (i.p.) injection of pH-adjusted saline or the corresponding antipsychotic drug for 28 consecutive days, as previously described (Barakauskas et al., 2010 (link)). Twenty-four hours after the last injection, all rats were sacrificed by decapitation, and brains were immediately removed and rinsed in ice-cold artificial cerebrospinal fluid. Frontal cortices were quickly dissected, frozen on dry ice, and stored at –80ºC. Tissue homogenization was performed as described above.