FIGURE 8.

miR-506 expression is elevated in cholangiocytes from patients with PBC. A, in situ hybridization of miR-506 expression using LNA-modified digoxigenin-labeled miR-506 specific or scrambled probes (as controls) was examined using formalin-fixed paraffin-embedded biopsies from normal livers and those of PBC patients. miR-506 was detected using cyanine 5 (red)-labeled peroxidase substrate following incubation with peroxidase-conjugated anti-digoxigenin antibodies. Cholangiocytes (bile ducts) were labeled by immunofluorescent staining using anti-rabbit IgG Alexa-488 (green) with a pan-cytokeratin primary antibody. Nuclei (blue) were labeled with DAPI. Scale bar (top right panel), 50 μm. Hybridization with the scrambled probe failed to show any significant cyanine 5 staining, suggesting specific binding of the miR-506 probe (data not shown). B, panel shows quantitation of the cyanine 5 pixel intensity/μm of bile duct in 5 control and 5 PBC liver sections. p < 0.05 using Mann-Whitney test.