VER-82576

Prior to treatments, the cells are placed into 6-well plates in medium at a density of 1 × 10⁵ cells/well, three wells per treatment group and cultured for 24 h. The cells are divided into treatment groups and treated accordingly for 40 h: vehicle control (DMSO, 0.016%, v/v), VER-82576 (0.05, 0.1 or 0.2 μM), irradiation (10 Gy), or VER-82576 (0.05, 0.1 or 0.2 μM) in combination with irradiation (10 Gy). Upon completion of the treatment, all cells are incubated with 0.5 mg/ml MTT for 3 h. The relative viability of the treated cells to the untreated control cells is measured. The absorbance is measured at 490 nm on a microplate reader^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

The estrogen receptor (ER)-positive, ErbB2-overexpressing cell line BT-474 is grown in DMEM high glucose (4.5 g/L) supplemented with 10% FCS, 200 mM l-glutamine, and 1% sodium pyruvate (BioConcept). Two or 3 d before cell inoculation, each mouse is s.c. implanted on the upper dorsal side with a 17β-estradiol pellet (25 μg/d; 90-d release) using a trocar needle. BT-474 cells (5 × 10⁶) are injected in 200 μL Matrigel/HBSS (1:1 volume) s.c. in the right flank. The A375 cell line expresses mutant B-Raf (V600E). The cells are grown in RPMI 1640 supplemented with 10% FCS and 200 mM l-glutamine. To establish tumors, 5 × 10⁶ A375 cells are injected s.c. in 200 μL PBS in the right flank. The animals are kept under optimized hygienic conditions with a 12-h dark, 12-h light cycle. The animals are fed food and water ad libitum. Tumor growth and body weights are monitored at regular intervals. The xenograft tumor sizes are measured manually with calipers, and the tumor volume is estimated using the following formula: (W × L × H × π/6), where width (W), length (L), and height (H) are the three largest diameters. Treatment with VER-82576 is initiated when the average tumor volume reaches 100 to 300 mm^3[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.