Mean DHE fluorescence was calculated by subtracting integrated density of the background signal from the integrated density of the fluorescent staining for 10 fields/heart, 5 hearts/group, and normalized to control.
After subtracting the average integrated density value of the corpus callosum from the integrated density values obtained from the cortical ROIs of that sample and analyzed the results for all four groups using a 2-way ANOVA.
The total fluorescence intensity of each cell was determined using ImageJ (integrated density, ID).
Data were expressed as integrated density volume (IDV) normalized to GAPDH.
All data were expressed as integrated density values (IDV).
The integrated density values were plotted against fraction number to profile component distribution.
The integrated density values (IDV) corresponding to the band intensities were measured.
Area, minimum and maximum gray value, integrated density, and mean gray value were measured.
Actual fluorescence quantification was recorded as integrated density on the entire combined tile image using Image J freeware (http://imagej.nih.gov/) using "integrated density", "mean value" and "area" as measurement settings.
For each image, the whole field was selected using the rectangle tool and measurements were set to take integrated density, area measured, and mean gray value (integrated density over area measured).
Organ measurements were obtained using the Freehand Selection tool to draw ROI around their perimeter and measuring integrated density.
