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1.
Figure 1.

Figure 1. From: Specificity, duplex degradation and subcellular localization of antagomirs.

Chemical modifications used in this study. Schematic representation of (A) miR-122, (B) antagomir-122, (C) antagomir-122 all P=S and (D) 5′-Quasar570 labeled antagomir-122.

Jan Krützfeldt, et al. Nucleic Acids Res. 2007 May;35(9):2885-2892.
2.
Figure 7.

Figure 7. From: Specificity, duplex degradation and subcellular localization of antagomirs.

Injection of antagomir-16 into mouse cortex. Northern blots of miR-16 and miR-124 from total RNA isolated from mouse cerebral cortex that had been injected with antagomir-16 or PBS into the right and left cerebral hemispheres, respectively. Each pair of lanes represents an individual animal.

Jan Krützfeldt, et al. Nucleic Acids Res. 2007 May;35(9):2885-2892.
3.
Figure 2.

Figure 2. From: Specificity, duplex degradation and subcellular localization of antagomirs.

Impact of antagomir phosphorothioate modifications and antagomir length on miR-122 levels. Northern blots of total RNA isolated from livers of mice that were treated with different antagomir-122 chemistries at 3 × 20 mg/kg bw. (A,B) Different phosphorothioate modifications; (C) different lengths. ‘P=S’ indicates phosphorothioate modification. Each lane represents an individual animal.

Jan Krützfeldt, et al. Nucleic Acids Res. 2007 May;35(9):2885-2892.
4.
Figure 4.

Figure 4. From: Specificity, duplex degradation and subcellular localization of antagomirs.

Sequence discrimination of antagomir-122. Steady-state mRNA levels of miR-122 target genes in livers of mice treated with the indicated amounts of antagomir-122 or antagomir-122 that harbored 4, 2 or 1 nt mismatches, respectively (A), or 1-nt mismatch at different positions (B). Expression was measured using RT-PCR. Gapdh was used as a loading control, Gapdh-RT denotes a control without reverse transcription. Each lane represents an individual animal.

Jan Krützfeldt, et al. Nucleic Acids Res. 2007 May;35(9):2885-2892.
5.
Figure 6.

Figure 6. From: Specificity, duplex degradation and subcellular localization of antagomirs.

Localization of antagomir-122 and miR-122 in hepatocytes. Liver tissue from mice that were treated with 3 × 80 mg/kg Q570-labeled mm-antagomir-122 was fractionated on a sucrose gradient following ultracentrifugation. Localization of Q570-labeled mm-antagomir-122 was analyzed by spectrophotometry (A), localization of t-RNA and miR-122 were analyzed using northern blotting of total RNA isolated from each fraction (B). For subcellular localization of antagomirs and P-bodies in mouse liver, mice were treated with Q570-labeled antagomir-122 and a DNA-plasmid expressing a GFP-GW182 hybrid as described in the Materials and Methods section. P-body and Q570-antagomir localizations were visualized using laser-scanning microscopy (C).

Jan Krützfeldt, et al. Nucleic Acids Res. 2007 May;35(9):2885-2892.
6.
Figure 3.

Figure 3. From: Specificity, duplex degradation and subcellular localization of antagomirs.

Dose- and time-dependency of miR-122 target regulation by antagomir-122. Steady-state mRNA levels of miR-122 target genes in livers of mice treated with the indicated amounts of antagomir-122. Expression was measured by RT-PCR. Each lane indicates an individual animal. The glyceraldehyde-3-phosphate dehydrogenase gene (Gapdh) was used as a loading control. Gapdh-RT denotes a control without reverse transcription. Gene symbols are shown in accordance with the International Standardized Nomenclature (www.informatics.jax.org/mgihome/nomen/gene.shtml). The upper row shows a northern blot of liver RNA for miR-122. Each lane represents an individual animal. (A) Dose-dependency. (B) Time-course.

Jan Krützfeldt, et al. Nucleic Acids Res. 2007 May;35(9):2885-2892.
7.
Figure 5.

Figure 5. From: Specificity, duplex degradation and subcellular localization of antagomirs.

Regulation of miR-122 targets by chemically protected antagomir-122/miR-122-duplexes. (A) Schematic description of the two different duplexes used. (B) Steady-state mRNA levels of miR-122 target genes in livers of mice treated with the indicated modified antagomir-122/miR-122-duplexes. Expression was measured using RT-PCR. Fold-regulation indicates the ratio of expression levels of the means of mice treated with antagomir-122/miR-122 duplex compared to the PBS group. The upper row shows a northern blot of liver RNA for miR-122. As controls, duplexes were added to 5 μg total kidney RNA and loaded on polyacrylamide gels before (‘input’) or after the Trizol protocol (‘Trizol’). Each lane represents an individual animal. Gapdh was used as a loading control, Gapdh-RT denotes a control without reverse transcription. *P < 0.05; **P < 0.01; ***P < 0.001; Student's t-test compared to PBS.

Jan Krützfeldt, et al. Nucleic Acids Res. 2007 May;35(9):2885-2892.

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