Each ring was placed in a collagen pre coated 96 well plate. VEGF, with or without different dilutions of santalol or selleck chem Lenalidomide sunitinib, was added to the wells. On day 6, the rings were analyzed by phase contrast microscopy and microvessel outgrowths were quantified and photographed. The assay was scored from 0 to 5 in a double blind Inhibitors,Modulators,Libraries manner. Each data point was assayed 6 times. Sponge implant angiogenesis assay Sponge implant assay was performed as described previ ously. Sterile circular sponge discs were inserted subcutaneously into male Swiss albino mice. The day of sponge insertion was taken as day 0. Commencing day 1, animals were treated with santalol from day 1 to day 14. On the day following Inhibitors,Modulators,Libraries the last injection, the sponges were excised, photographed and weighed. Sponges were bisected.

one half was fixed in 10% formalin and embedded in paraffin wax. Sections were stained Inhibitors,Modulators,Libraries with hematoxylin/eosin for identification of blood vessels. Immunostaining was done for VEGF and CD31. The second half of the sponge was weighed, homogenized in 2 ml of sterile PBS at 4 C, and centrifuged to quantify level of VEGF. The VEGF in the supernatant from each implant were measured in 50 ul of the supernatant using Immuno assay Kits following the manufac turers protocol. The extent of the vascularization of the sponge implants was assessed by the amount of Hemoglobin detected in the tissue using the Drabkin method. All procedures for animal experimentation used were approved by the Institutional Animal Ethics Com mittee, King Saud University, Riyadh, Saudi Arabia.

Xenograft human prostate tumor mouse model Six week old male BALB/cA nude mice were purchased Inhibitors,Modulators,Libraries from Charles River Laboratories. Animals were housed in a specific pathogen free room within the animal facilities at the King Saud University, Riyadh. All animals were allowed to acclimatize to their new envir onment for one week prior to use and were handled ac cording to the Institutional Animal Care and Use, King Saud University, Riyadh. Mice were randomly divided into 3 groups. PC 3 cells were resuspended in serum free RPMI1640 medium with matrigel basement membrane matrix at a 1 1 ratio and then were subcutaneously injected into the flanks of nude mice. After tumors grew to about 100 mm3, mice were treated intraperitoneally with or without santalol daily for 15 days. 0. 1% DMSO served as ve hicle control.

The body weight of each mouse was re corded and tumor volume was determined by Vernier caliper Inhibitors,Modulators,Libraries every day, following the formula of A B2 0. 52, where A is the longest diameter of tumor and B is the shortest diameter. After 16 d, the mice were killed by cervical dislocation and solid tumors were removed. Survival was evaluated http://www.selleckchem.com/products/BI6727-Volasertib.html by the Kaplan Meier method. Mice of each group were also monitored for other symp toms of side effects including food and water withdrawal and impaired posture or movement.