Figure 8.

Repression of expression of selected candidate target genes of miR-34a and miR-34c by transfection analysis. Human embryonic kidney (HEK 293) cultures were transfected with green fluorescence protein (GFP) expression vectors containing sequences encoding miR-34a, miR-34c, or a scrambled sequence. Controls included un-infected cultures, mock transfected cultures, cells transfected with either GFP vector alone or carrying a computer-generated scrambled sequence. (A) Western blot analysis of cell lysates probed for SIRT1, Onecut2, Presenilin-1, Bcl2, and β-actin proteins; (B) Graphic representation of a comparison of band intensities between the various treatments; image intensities of Western blotted bands were normalized against the β-actin band, which is constant throughout. Graphic representation showing percent of repression, estimated using scrambled controls for comparison (panels C–E): transfection with miR-34a (C); miR-34c (D); miR-34a and miR-34c (E). One-way ANOVA followed by LSD test was used to determine statistical significance; ^*P < 0.05, ^**P < 0.01.