Development of MIR17PTi. (A) Illustration summarizing the activity of LNA gapmeRs with regard to the miR-17-92 cluster. (B) qRT-PCR analysis of pri-mir-17-92 expression in 293T 2 days after transfection with miR-17-92 LNA gapmeRs or scr-NC (25 nM). (C) qRT-PCR analysis of miR-17-92s in 293T 2 days after transfection with mir-17-92 LNA gapmeRs or scr-NC (25 nM). (D) qRT-PCR analysis of pri-mir-17-92 expression in 293T 2 days after transfection with mix-MIR17PTi or scr-NC (25 nM). (E) qRT-PCR analysis of pri-mir-17-92 (left) and western blotting of RNase H1 (right) in 293T cotransfected with either small interfering RNAs (siRNAs) targeting RNase H1 (siRNASE H1; 25 nM) or scrambled siRNAs (siCNT; 25 nM) and MIR17PTi or scr-NC (25 nM; 3-day time point). GAPDH was used as protein loading control. (F) qRT-PCR analysis of pri-mir-17-92 in 293T during a time-course exposure (days 1.5, 3, 4.5, and 6, every 36 hours) to MIR17PTi or scr-NC (10 µM). (G) Confocal microscopy analysis of 293T after 3 or 4.5 days of exposure to FAM-labeled MIR17PTi (10 µM). Cell nuclei are evidenced by 4′,6-diamidino-2-phenylindole staining (original magnification ×40). (H) qRT-PCR analysis of miR-17-92s in 293T during a time-course exposure (days 1.5, 3, 4.5, and 6, every 36 hours) to MIR17PTi or scr-NC (10 µM). The qRT-PCR results are average expression levels after normalization with GAPDH (for pri-mir-17-92) or RNU44 (for miR-17-92s) and ΔΔCt calculations. Data from 1 of 3 independent experiments are shown in each panel. *P < .05.