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. 2015 Oct 16;6(35):37852–37870. doi: 10.18632/oncotarget.5635

Figure 3. miR-451 MRE-regulated adenovirus specifically delivers lincRNA-p21 into CRC cells and CSCs.

Figure 3

A. The expression levels of miR-451 in normal mucosa and CRC samples of distinct grades of differentiation (Inter, intermediate; undif, undifferentiated) were measured by qPCR. U6 was used as an endogenous control. B. The levels of miR-451 were quantified by qPCR in normal cell lines and ALDH+ or ALDH− subsets of CRC cells. U6 was used as endogenous control. C. Luciferase activity was detected in indicated types of cells after transfection with psiCheck (1 μg / 1 × 106 cells) and psiCheck2-miR-451-MRE (1 μg / 1 × 106 cells) for 48 hrs. D. Four copies of miR-451 MREs were inserted immediately following a lincRNA-p21-coding ORF on adenoviral vector Ad-lnc-p21 to generate Ad-nc-p21-MRE. Ad-EGFP served as control. E. The cells were infected with Ad-EGFP, Ad-lnc-p21, or Ad-lnc-p21-MRE at 10 MOI for 48 hrs. LincRNA-p21 expression was detected by Northern blotting. U6 was used as an endogenous control. F. LincRNA-p21 was further quantified by qPCR in these cells. U6 was a loading control. Representative images (E) are shown. Data are presented as the mean ± SD (A, B, C, F) of each group from triple replicates. *P < 0.05, **P < 0.01.