Figure 6.

Limited in vivo effects of RO1

Tumors were generated by orthotopic injection of 100,000 ZH-161 or T-325 cells in nude mice. Beginning on day 14 after implantation, mice were treated daily for 7 days by oral gavage with RO1 (30 mg/kg in 0.3% Tween 20 in saline) or solvent control. Three animals per group were euthanized on day 21 after tumor cell implantation for tissue analyses. (A) Tumor volumes were determined by area measurement on at least five hematoxylin and eosin (H&E)-stained slides per tumor utilizing ImageJ followed by calculation per stack height. (B) Survival was defined as the time from tumor cell implantation to the development of neurological symptoms or loss of at least 15% weight (N = 5–7 mice per group, log rank test not significant). (C) Tumor invasion was determined by counting of tumor cell islands, defined as at least 50 cells at a distance of at least two cell layers from the tumor bulk (yellow lines). (D) Apoptosis was determined by cleaved caspase-3 staining (arrowheads indicate groups of caspase-3-positive cells).