All animal protocols described were approved by the Institutional Animal Care and Use Committee at The University of Arizona in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Four-week female athymic nude mice were purchased from Harlan Laboratories and housed in plastic cages in laminar flow isolated hoods with access to water and autoclaved mouse chow ad libitum. Mice were inoculated in the left cardiac ventricle with MDA-MB-231 tumor cells as previously described.5 (link) Briefly, tumor cell inoculation was performed percutaneously into the left cardiac ventricle on anesthetized mice in a supine position with a 26-gauge needle attached to a 1 mL syringe containing 1 × 105 cells suspended in 0.1 mL sterile PBS. Visualization of bright red blood entering the hub of the needle in pulsatile fashion was indicative of correct needle placement into the left cardiac ventricle. After tumor inoculation, mice were divided randomly into three groups (n = 11), treated by intraperitoneal (ip) injection with 25 or 50 mg/kg curcuminoids or vehicle alone every other day beginning on day 1 for 21 days and followed prospectively for the development of bone lesions by radiography on days 12, 16 and 21 of the experiment. Mice were X-rayed in a prone position using a Digital Faxitron MS-20 with a digital camera at 4x magnification. Lytic lesion area, reported as total lesion area per animal in the hind limb long bones, was analyzed in a blinded fashion by three independent investigators using ImageJ 1.43u software (National Institutes of Health).