DSS-induced alterations in goblet cells and subsequent depletion in synthesis and secretion of mucin glycoprotein (MUC2) was analysed by Alcian blue staining (ab150662 Alcian Blue, pH 2.5 (Mucin Stain), Abcam, Australia) following the manufacturer’s instructions. Briefly, paraffin-embedded colon sections (n = 4/group) were stained with Alcian blue, staining the acidic sulphated mucin blue and the counterstained with Safranin O, staining the nuclei red following the method previously described [30 (link)]. Computer-assisted image analysis was performed with a Leica DM500 microscope (Leica Microsystems, Wet-zlar, Germany) and Leica ICC50 W camera (Leica Microsys-tems, Wetzlar, Germany). The staining intensity (IOD) was assessed using Image Pro Plus 7.0 (Media Cybernetics, Inc., Rockville, MD, USA) and used for comparison among groups [31 (link)].
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