Fig. 4

HDAC inhibition and sample preparation influence lectin binding in fixed cells. SW13 cells were left untreated (Control), or were treated with 1 nM FK228 for 24 h (FK228). Cells were then fixed and left unpermeabilized (a), or were permeabilized with 0.2% Triton-X (b) before incubation with 20 μg/ml FITC labeled lectins for ~ 2 h. To demonstrate specificity of lectin binding, wheat germ agglutinin (WGA) stained cells were incubated with WGA elution buffer (+ Comp) for 30 min. Permeabilization of fixed cells before incubation with FITC-labeled lectins can significantly impacts staining of some lectins. All samples were mounted with ProLong Gold anti-fade reagent with DAPI to stain nuclei and photographed using a Zeiss Axiovert apotome with a 40X objective lens and a uniform exposure at each wavelength