Fig. 3

Comparison of analysis by IOCBIO Gel and ImageJ Gel plugin. A A representative example of a Western blot membrane labeled with primary and secondary antibodies to assess GAPDH expression (image acquired as a part of [16]). Insets a–d illustrate bands that are either easy to analyze due to clear separation between the lanes (b, c) and that are more complicated due to overlap with the ladder on the left (a) or on the right (d). B Results of the expression estimation performed by three users using either IOCBIO Gel or ImageJ Gels plugin to analyze the image shown in A. Notice the very small spread of the estimations in the most of the lanes and the larger spread on the first (shown in inset a of A) and the two last lanes (inset d in A corresponds to the last lane). C Expression of GAPDH, L-type calcium channel (LTCC), and estimation of protein loading using the Ponceau staining performed by IOCBIO Gel and ImageJ Gels plugin done on three separate membranes and by three different users (analyzed images acquired as a part of [16]). For comparison, the diagonal line corresponding to the equality in estimation is shown in black. Each dot on the plots corresponds to two estimations performed by specific user (shown by color) for the same lane on a gel using IOCBIO Gel (y-axis) and ImageJ Gels plugin (x-axis). Notice that the dots are distributed along the diagonal line with only small, occasional deviations. This demonstrates that the estimations by both software solutions are the same