2.3.1. Animals

A total of 50 specific pathogen-free C57BL/6 mice (males, 25–30 g) were obtained and maintained at the animal facilities of the Gonçalo Moniz Institute-FIOCRUZ (Salvador, BA, Brazil). Animals were housed in cages with free access to food and water. All animals were kept under a 12:12 h light–dark cycle (lights on at 6:00 a.m.). The animals were treated according to the ethical principles for animal experimentation of SBCAL (Brazilian Association of Laboratory Animal Science), Brazil. The Animal Ethics Committee of the Oswaldo Cruz Foundation (Salvador, BA, Brazil) approved the experimental protocol (number 01/2013).

#### 2.3.2. In Vivo B16-F10 Melanoma Model

The in vivo antitumor effect was evaluated in C57BL/6 mice inoculated with B16-F10 melanoma cells, as previously described in ref. [37]. Tumor cells (2 × 106 cells per 500 <sup>μ</sup>L) were implanted subcutaneously into the left hind groin of mice. The compounds were dissolved in 5% DMSO and provided to the mice intraperitoneally once a day for 15 consecutive days. Mice were divided into five groups at the beginning of the experiment: Group 1 (negative control, *n* = 10): animals treated with vehicle (5% DMSO); Group 2 (positive control, *n* = 10): animals treated with DOX (1 mg/kg/day); Group 3: animals treated with CUR (20 mg/kg/day, *n* = 10); Group 4: animals treated with DSS (60 mg/kg/day, *n* = 10); Group 5: animals treated with CUR (20 mg/kg/day) plus DSS (60 mg/kg/day) (*n* = 10). The treatments were initiated one day after tumor injection. On day 16, the animals were anesthetized (thiopental, 50 mg/kg), and samples of peripheral blood were collected from the brachial artery for hematological analyses, as described below. The animals were euthanized by anesthetic overdose (thiopental, 100 mg/kg), and the tumors were excised and weighed. Drug effects are expressed as the percentage inhibition in relation to the control.
