Figure 3.

Subcellular localization of MAH. 21PT epithelial cells grown on coverslips were fixed by paraformaldehyde, then permeabilized with 0.5% Triton X-100 (a and a′) or permeabilized twice, first by 0.05% Triton X-100 before fixation, then after fixation as above (b and b′). After double permealization cells were treated with DNase I (c and c′) or DNAse I and RNase A (d and d′). The fixed samples were incubated with a 1:250 dilution of C12 antiserum, and subsequently with rhodamine-conjugated secondary antibody (a–d) followed by counterstaining with the DNA dye DAPI (a′–d′) before analysis with confocal laser beam microscopy.