Tetrathiomolybdate ttm

5 × 10⁶ MEFs or 10⁷ melanoma cells resuspended in phosphate buffer saline were injected subcutaneously into flanks of SCID/beige mice (Charles River Laboratory) as previously described^{42 (link)}. Drug treatments were as follows: vehicle [1% methylcellulose (Sigma), 1% dimethyl sulfoxide (DMSO, Sigma)], 2.0 mg tetrathiomolybdate (TTM, Sigma) in vehicle, or 20 mg/kg vemurafenib (Chemietek) in vehicle every other day (q.o.d) via oral gavage. Mice were fed a normal diet (PMI 5053 Picolab Mouse Diet 20, LabDiet) or where indicated, a Cu-deficient diet (CuD diet, TD.80388, Harlan Teklad). Mice fed a CuD diet mice were administered deionized H₂O (diH₂O) supplemented with 20 mg/L CuSO₄, diH₂O alone, or diH₂O supplemented with 3 g/L trientine dihydrocholoride (Sigma). All studies were approved by the Duke University Institutional Animal Care and Use Committee. Statistical analysis of tumor volumes at end point was performed using a one-tailed, unpaired T-test, with a 95% confidence interval for two group datasets or one-way analysis of variance (ANOVA) with a 95%, 99%, or 99.9% confidence interval and Tukey’s multiple comparison post test for ≥ 3 datasets in Prism 5 (GraphPad). Statistical analysis of percentage of mice with tumors ≥ 1.0 cm³ versus time (days) was analyzed using a survival curve log-rank (mantel-cox) test in Prism5 (GraphPad).