Figure 2

In vivo testosterone-HSA effects on tumor incidence in APC^Min/+ mice. APC^Min/+ mutant mice of both sexes were used in this study. (A) Confocal laser scanning microscopic analysis of APC^Min/+ colon tumor frozen sections stained with testosterone-HSA-FITC or HSA-FITC, showing specific FITC-related fluorescence at the cell membranes. Visualization of the nuclei was evident by DRAQ5™ staining. Magnification 100×. (B) Arithmetic means ± SEM of colonic tumor incidence in APC^Min/+ mice. The mice were either treated with 5 mg/kg testosterone-HSA subcutaneously 3 times/wk for 8 wk, (n = 6 animals, black bar) or treated with 5 mg/kg of normal saline subcutaneously 3 times/wk for 8 wk (n = 4 animals, blue bar); ** indicates significant difference between both groups (P < 0.01). (C) After treatment, the ApC^Min/+ colonic cancer tissue was cut to 8-µm frozen sections, and fragmented DNA was assessed using TUNEL assay according to the manufacturer’s instructions. Representative confocal laser scanning microscopy analyzed samples from at least three independent tumors from each animal; magnification 100×. (D) Representative confocal laser scanning microscopic analysis of TAC-treated and untreated APC^Min/+ frozen colon tumor sections prepared from at least three tumors of similar size from each animal stained with (a) anti-phospho-Akt (Thr308) and (b) anti-phospho-Bad (Ser136) antibodies. Antirabbit FITC was used as secondary antibody and DRAQ5™ for nuclei staining; magnification 100×.