Cremophor el ethanol

Female BALB/c mice, 6–8 weeks of age, were obtained from NCI–Frederick (Frederick, MD, USA). BNL, a murine HCC cell line, was kindly provided from University of Navarra, Pamplona Spain^{11 (link)}. Mycoplasma testing was performed by SAIC Frederick two months prior to experiments Cells were routinely kept in culture for no more than 8 to 10 passages. Mice were injected s.c. with 1 x10⁶ BNL cells. One week after tumor inoculation when tumors are palpable, mice received a daily oral gavage of sorafenib (Bayer) at a dose of 10 mg/kg. sorafenib stock solution (4x) was freshly prepared every 4 days using Cremophor EL/ethanol (50:50; Sigma). The final 1x dosing concentration was prepared by diluting with sterile water immediately prior to administration to mice. Control mice received vehicle. Endoglin antibody (clone MJ7/18), which binds murine endoglin, was purchased from the Developmental Studies Hybridoma Bank at the University of Iowa and purified at NCI. 100 mg/mouse was given i.p. every other day. Tumors were measured every other day using digital calipers. All mice were handled, fed, and housed in accordance with the U.S. Department of Health and Human Services institutional guidelines.
Experimental protocol was approved by NCI Animal Care and Use Committee (ACUC).