Confirmation of galectin-3 expression at the protein level using western blotting and immunohistochemistry. Western blotting was performed at 28 days and 6 months post-injury, and showed a significant increase in expression in moderately injured spinal cord over sham-injured tissue. Representative western blots and graphical representations are shown in A, B. Immunohistochemistry was performed at 28 days post-injury. Sham-injured tissue demonstrated no positive immunostaining for galectin-3 (C, D). Galectin-3 positive cells (labeled with a fluorescent red secondary antibody) were found throughout injured tissue at this time point, and labeled large, ameboid cells that are typical of macrophages and activated microglia (E, F). DAPI (blue) was used as a counterstain for cells. Insets show magnified areas from the white matter. Injured tissue double-labeled for microglia/macrophages (Iba-1) and galectin-3 (arrows) demonstrate correlation in expression in the merged image (G). Size bar = 500 μM (C, E); 100 μM (D, F); 75 μm (G). Bars represent mean +/- SEM.