Figure 4. Flanking exons or deletion of... | Rockefeller University Press

$Figure 4. Flanking exons or deletion of the CPA signal results in increased levels of an intronic pri-miRNA. (A) Schematic of pri-miR-26b constructs. Flanking exons (E) are represented as blue rectangles. Probes used for ISH are shown as blue lines under pri-miR-26bFE. (B, top and middle) Northern blotting was performed on cells transfected with constructs in A as described in Fig 1. RT, readthrough transcripts. (B, bottom) Mature miRNA levels were quantitated and normalized to the U6 snRNA, and the amount of miR-26b produced from pri-miR-26b was set to 1. Error bars represent standard deviations (n = 4). (C) Nuclear fractionation of released and chromatin-associated transcripts. Cells transfected as in B were fractionated as described in Fig 3. P, pellet; S, supernatant. (D) ISH of pri-miR-26b transcripts in cells transfected as in B using DIG-labeled probes (Fig. 4 A, blue lines). Nuclei were stained with DAPI and merged images are shown in panels e–h. Bar, 10 μm.$

Figure 4.

Flanking exons or deletion of the CPA signal results in increased levels of an intronic pri-miRNA. (A) Schematic of pri-miR-26b constructs. Flanking exons (E) are represented as blue rectangles. Probes used for ISH are shown as blue lines under pri-miR-26bFE. (B, top and middle) Northern blotting was performed on cells transfected with constructs in A as described in Fig 1. RT, readthrough transcripts. (B, bottom) Mature miRNA levels were quantitated and normalized to the U6 snRNA, and the amount of miR-26b produced from pri-miR-26b was set to 1. Error bars represent standard deviations (n = 4). (C) Nuclear fractionation of released and chromatin-associated transcripts. Cells transfected as in B were fractionated as described in Fig 3. P, pellet; S, supernatant. (D) ISH of pri-miR-26b transcripts in cells transfected as in B using DIG-labeled probes (Fig. 4 A, blue lines). Nuclei were stained with DAPI and merged images are shown in panels e–h. Bar, 10 μm.

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