Wheat Germ Agglutinin (WGA) staining

Paraffin-embedded left ventricle tissue sections (4 μm in thickness) were deparaffinized and rehydrated in four changes of Xylene, 2 changes of 100% ethanol, 2 changes of 95% ethanol, and 2 changes of 70% ethanol. The tissue sections were then rinsed with tap water and incubated with Oregon Green 488 WGA solution (5 μg/ml) from Life Technologies (Cat. No. W6748) at 4°C overnight. Afterwards, slides were washed three times with PBS. The slides were allowed to air dry, and then mounted with Prolong Gold Antifade Reagent from Life Technologies (Cat. No.: {"type":"entrez-protein","attrs":{"text":"P36930","term_id":"1248281091","term_text":"P36930"}}P36930). Eight fluorescent images were randomly taken from each tissue section using an Olympus fluorescent microscope with a 20x lens. From each image, the cross sectional area of 20 cells was measured using ImageJ as previously described [35].