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. Author manuscript; available in PMC: 2013 Feb 1.
Published in final edited form as: Hepatology. 2011 Dec 19;55(2):465–475. doi: 10.1002/hep.24698

Figure 6. DR4 Repression by miR-25 and Functional Rescue Via miR-25-resistant DR4.

Figure 6

Panel A: After transfection of KMCH cells with control locked nucleic acid (CT) or LNA against miR-25 (LNA-25) for 48 hours, slides were prepared and analyzed by confocal microscopy for immunofluorescence intensity of DR4 or DR5. Panel B: Fluorescence intensity of cells transfected and stained as in panel A was quantified by ImageJ software. The data are presented as fold change in the average DR4 or DR5 intensity, mean +/-SEM, ***p<0.001. Panel C: H69 cells were transfected (24hr) with control (CT) or mir-25 precursor; slides were prepared and examined for DR4 immunofluorescence by confocal microscopy. Panel D: DR4 fluorescence intensity in H69 cells was quantified by ImageJ software and presented as fold change in average signal intensity between control and treatment groups, mean +/- SEM, ***p<0.001. Panel E: H69 cells were transfected with GFP plus pCDNA, GFP plus pCDNA-mir-25, or DR4-GFP plus pCDNA-mir-25. Eighteen hours after transfection, TRAIL was added at 4 ng/ml for additional 6 hours to induce apoptosis. Cells were stained with DAPI and GFP-positive cells were evaluated based on nuclear morphology for presence of apoptosis. Data are presented as the percentage of cells with apoptotic nuclear morphology out of total cell count (mean +/-SEM, **p<0.01, ***p<0.001).