Additional file 2: Figure S2. of Enhancement of the CXCL12/CXCR4 axis due to acquisition of gemcitabine resistance in pancreatic cancer: effect of CXCR4 antagonists
Quantification of immunostaining of CXCR4 and CXCL12 protein by digital image analysis. (A) The number of CXCR4 immunoreactive cells in mouse specimens was expressed as a percentage of the total number of cells that were randomly counted in 10 fields at × 400 magnification. Furthermore, for each image, the color deconvolution method was used to isolate CXCL12-positive DAB-stained cells from CXCL12-negative hematoxylin-stained cells. The measurement parameter was IOD. Optical density was calibrated and the area of interest was set as follows: hue, 0–30; saturation, 0–255; intensity, 0–255. (B) The values were determined, and the IOD was log10 transformed. Values are expressed as means ± SD. Multiple comparisons were performed using one-way ANOVA followed by Bonferroni test, **, P < 0.01; *, P < 0.05. (JPG 295 kb
