Increased expression of PCNA, NE, and pAKT^Ser473.

<p>(<b>A</b>) PCNA⁺ cells were detected in the colon tissues. The colons were removed from the AOM and DSS-treated mice at the indicated times. Immunohistochemical analysis was performed using anti-PCNA antibodies as described in the Materials and Methods. The numbers of cells per field were counted in 5 randomly selected visual fields at 400× magnification. Values represent mean ± SD for 7 mice in each group. *p<0.05, **p<0.01 versus day 0 mice. (<b>B</b>) <i>NE</i> mRNA expression in the colons of the two groups sacrificed at the indicated times was assessed by quantitative RT-PCR. Each value represents mean ± SD (n = 7 mice/group). *p<0.05, **p<0.01 versus control mice or day 0 mice. (<b>C</b>) Immunoblotting analysis with anti-phospho-Akt (p-Akt) and anti-Akt (Akt) antibodies was performed using cell lysates of the colon tissues of the AOM and DSS-treated (tumor) and control (con) mice sacrificed on days 28 and 35, as described in the Materials and Methods. Representative results of 3 independent experiments are shown. β-actin served as a control for the mRNA levels. (<b>D</b>) Neutrophils expressing NE were detected by double-color immunofluorescence analyses using anti-Ly6G (red) and anti-NE (green) antibodies, as described in the Materials and Methods. The fluorescence images are digitally merged in the far right image. Representative results of 7 mice are shown. Original magnification, 630×.