Immunohistochemical staining for intratumor caspase-3⁺ apoptotic cells.

<p>Tumor tissue sections were prepared after treatment with saline (panel <b><i>A</i></b>), manuka honey (panel <b><i>B</i></b>), taxol (panel <b><i>C</i></b>) or manuka+taxol (panel <b><i>D</i></b>) and stained using caspase 3-specific antibody, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055993#s2" target="_blank">Materials and Methods</a>. Representative images at high magnification (bar = 50 µm) are shown. Arrows indicate representative, brown-staining, apoptotic cells. Necrotic regions are also indicated (n). The results are representative of two independent experiments. (<b><i>E</i></b>) Quantitative estimation of the number of caspase-3 positive cells in tumor sections of different treatment groups. The data is shown as the mean ± SEM of the number of positive cells per high power field. Tumors were obtained from 2–3 mice per treatment group and multiple sections were made from each tumor tissue. The number of positive cells was determined by counting the number of cells in 20 high power fields per section. Asterisks denote statistically significant differences between each experimental group and the saline control group; also shown is a comparison between manuka alone and manuka+taxol groups (*, <i>p</i><0.05).