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Results: 6

1.
Fig. 1.

Fig. 1. From: MicroRNA-206 colocalizes with ribosome-rich regions in both the nucleolus and cytoplasm of rat myogenic cells.

In situ hybridization pattern of miR-206 compared with let-7 and SRP RNA. (Upper) Distribution of signal after in situ hybridization using LNA probes to miR-206, let-7, and SRP RNA. Images are not scaled to the same intensity ranges. All probes were cy3-labeled except let-7, which was fluorescein-labeled; miR-206 and let-7 patterns are shown after a dual hybridization experiment. (Lower) The corresponding phase images for miR-206 and let-7. (Lower Right) The hybridization pattern to SRP RNA using a peptide nucleic acid probe. PNA, peptide nucleic acid. Arrows point to the nucleolus. Each image is 35 μm wide.

Joan C. Ritland Politz, et al. Proc Natl Acad Sci U S A. 2006 December 12;103(50):18957-18962.
2.
Fig. 6.

Fig. 6. From: MicroRNA-206 colocalizes with ribosome-rich regions in both the nucleolus and cytoplasm of rat myogenic cells.

miR-206 partially colocalizes with 28S rRNA in the nucleolar GC and the cytoplasm. AF and GL depict results from two different cells. A and G show the miR-206 signal in a deconvolved mid-plane of the cell, and C and I show the 28S rRNA signal in the same plane. B and H are pseudocolored images combined to reveal overlap. D and J show enlarged images of the nucleoli. E and K are intensity linescans along the white line in D and J, respectively. F and L are intensity linescans along the white line in B and H, respectively. Images are 35 μm wide (AC and GI), 5.8 μm wide (D), and 4.1 μm wide (J).

Joan C. Ritland Politz, et al. Proc Natl Acad Sci U S A. 2006 December 12;103(50):18957-18962.
3.
Fig. 5.

Fig. 5. From: MicroRNA-206 colocalizes with ribosome-rich regions in both the nucleolus and cytoplasm of rat myogenic cells.

miR-206 is not concentrated in nucleolar FCs but is concentrated in the GC. L6 myoblasts were subjected to immunostaining for UBF (to mark FCs, AH) or nucleostemin (to mark the GC, IP) followed by in situ hybridization to miR-206, and image stacks were captured and subjected to 3D deconvolution. Enlarged images of deconvolved nucleoli show miR-206 (A and D) and corresponding UBF (C and F) distribution, and pseudocolored images (B and E) show miR-206 (red) and UBF (green) overlap (yellow). Linescan in G corresponds to the line in B, and linescan in H corresponds to the line in E. Similarly, enlarged images of deconvolved nucleoli show miR-206 (I and L) and corresponding nucleostemin (K and N) distribution, and pseudocolored images (J and M) show miR-206 (red) and nucleostemin (green) overlap (yellow). Linescan in O corresponds to the line in J, and linescan in P corresponds to the line in N. Images are 9.9 μm wide (AC), 5.9 μm wide (DF), 7 μm wide (IK), and 7.3 μm wide (LN).

Joan C. Ritland Politz, et al. Proc Natl Acad Sci U S A. 2006 December 12;103(50):18957-18962.
4.
Fig. 4.

Fig. 4. From: MicroRNA-206 colocalizes with ribosome-rich regions in both the nucleolus and cytoplasm of rat myogenic cells.

miR-206 is not concentrated in nucleolar DFC. L6 myoblasts were transfected with a plasmid encoding GFP-fibrillarin (AE) or subjected to immunostaining for fibrillarin (FJ) followed by in situ hybridization for miR-206, and image stacks were captured and subjected to 3D deconvolution. A and B show raw images of miR-206 and fibrillarin-GFP signal. C and E show the corresponding deconvolved nucleoli. D shows a pseudocolor image of C and E overlap where miR-206 is red and fibrillarin is green. Similarly, F and G show raw images of miR-206 and fibrillarin signal after antibody staining and in situ hybridization, H and J show the corresponding deconvolved nucleoli, and I shows a pseudocolor image of H and J overlap. Yellow indicates areas of minimal overlap between fibrillarin and miR-206. (K) Linescan showing intensity of both miR-206 (red) and fibrillarin-GFP (green) along the line shown in D. (L) Linescan showing intensity of both miR-206 (red) and antibody-detected fibrillarin (green) along the line shown in I. Images are 35 μm wide (A, B, F, and G), 5.7 μm wide (CE), and 6.8 μm wide (HL).

Joan C. Ritland Politz, et al. Proc Natl Acad Sci U S A. 2006 December 12;103(50):18957-18962.
5.
Fig. 2.

Fig. 2. From: MicroRNA-206 colocalizes with ribosome-rich regions in both the nucleolus and cytoplasm of rat myogenic cells.

miR-206 increases during myogenesis. (A) Histogram showing relative levels of miR-206 in L6 nucleoli, nucleoplasm, and cytoplasm during myogenesis. (B) Same as A, but a scrambled control probe was used for hybridization. Similarly sized regions in each compartment were circled, and the average intensity per pixel was recorded. The averages of these measurements in multiple cells are shown here. It should be noted that this experiment has a low signal-to-noise ratio compared with the other experiments presented in this article because it was necessary to use confluent cultures to induce differentiation. In situ hybridization signal is substantially reduced in confluent cell cultures because of permeability issues. Bars indicate standard errors.

Joan C. Ritland Politz, et al. Proc Natl Acad Sci U S A. 2006 December 12;103(50):18957-18962.
6.
Fig. 3.

Fig. 3. From: MicroRNA-206 colocalizes with ribosome-rich regions in both the nucleolus and cytoplasm of rat myogenic cells.

Mature miR-206 probe hybridizes in a different pattern than a probe to premiR-206, and hybridization is RNase-sensitive. Dual color in situ hybridization was performed by using a cy3-labeled LNA probe to miR-206 (A) and a fluorescein-labeled LNA probe to premiR-206 (B). (C) Phase contrast image. Red arrows point to the nucleoli. (D and E) Control using cy3-labeled LNA miR-206 probe alone. (D) Red channel. (E) Green channel. This image shows a nucleus where the nucleolar:nucleoplasmic ratio of miR-206 is ≈1:1. (F) Cy3-labeled scrambled LNA probe. A, D, and F are scaled the same, and B and E are scaled the same. (G) Fixed cells were incubated with RNase buffer alone before in situ hybridization with cy3-labeled miR-206 LNA probe. (H) Cells were treated with a mixture of three ribonucleases (see Materials and Methods) before in situ hybridization with cy3-labeled miR-206 LNA probe. G and H are scaled the same. All images are 35 μm wide.

Joan C. Ritland Politz, et al. Proc Natl Acad Sci U S A. 2006 December 12;103(50):18957-18962.

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