A cohort of Tg-R266K Cbs−/− mice were studied under control (regular water), zinc water (25mM; 14 days), and zinc water supplemented with pyridoxine (0.4g/L; Sigma Aldrich P5669; 14 days) conditions. For most experiments only female mice were used as previous studies have shown that female mice more consistently induce Mt-I driven transgenes [Wang, et al., 2004 (link); Wang et al., 2005 (link)]. Mice ranged in age from 88–250 days of age. Blood was collected by retro-orbital bleed to measure serum tHcy and methionine.
Bortezomib (Velcade, Millennium Pharmaceuticals, Cambridge, MA, USA) was obtained from the pharmacy at Fox Chase Cancer Center. Adult female Tg-R266K Cbs−/− mice (n=8) were subcutaneously implanted with Alzet microosmotic pumps (Model 2001); pumping rate 1 microliter/hr) containing bortezomib diluted in 0.9% NaCl to deliver a final dose of 0.49 mg/kg/day. Mice were on zinc water 11 days before the pump implant and through the duration of drug studies, and were also eye bled before the implant to extract serum. Mice were sacrificed after two days on pump to harvest liver and serum.
Bortezomib (Velcade, Millennium Pharmaceuticals, Cambridge, MA, USA) was obtained from the pharmacy at Fox Chase Cancer Center. Adult female Tg-R266K Cbs−/− mice (n=8) were subcutaneously implanted with Alzet microosmotic pumps (Model 2001); pumping rate 1 microliter/hr) containing bortezomib diluted in 0.9% NaCl to deliver a final dose of 0.49 mg/kg/day. Mice were on zinc water 11 days before the pump implant and through the duration of drug studies, and were also eye bled before the implant to extract serum. Mice were sacrificed after two days on pump to harvest liver and serum.