Rat anti mac 2

Kidneys and other organs were obtained from WT and BTBR ob/ob mice after 14 weeks of drug therapy. Tissue sections were fixed, embedded, immunostained and analyzed as previously described (17 (link)), as were glomerular mesangial expansion and matrix accumulation. Briefly, the mesangial area occupied by silver methenamine stained matrix was quantified, as was the total area positive for matrix, by computer image analysis (Image Pro Plus, Media Cybernetics, Rockville, MD), for each glomerular cross-section. Mesangiolysis was determined by assessing the silver methenamine-stained glomeruli and scoring at least 50 glomeruli per section, scored present when lucency and dissolution of the normally compact silver-staining mesangial matrix was present and/or there were dilatation/ballooning of adjacent capillary loops.
Rat anti–Mac-2 (1:5000, Cedarlane, Hornby, Ontario, CA) and rabbit polyclonal anti-p57 (1:3000, Santa Cruz Biotechnology, Santa Cruz CA) were used to visualize glomerular macrophages and podocytes, as previously published (18 (link)). Quantification for Mac-2 and p57 were performed on digital images of immunostained tissues by counting, in a blinded manner, the number of Mac-2-positive cells and p57-positive podocytes per glomerular cross section in 50 glomeruli, per animal.