Mutational analysis of conserved positions comprising the C5–G57 base pair in the putative P1

<p><b>Copyright information:</b></p><p>Taken from "Core requirements for ribozyme self-cleavage reveal a putative pseudoknot structure"</p><p>Nucleic Acids Research 2006;34(3):968-975.</p><p>Published online 7 Feb 2006</p><p>PMCID:PMC1361622.</p><p>© The Author 2006. Published by Oxford University Press. All rights reserved</p>1 interaction. () Activity of six ribozyme construct designed to disrupt or maintain base pairing. Depicted is the proposed P1.1 interaction alone for each construct in the same orientation as shown in , where mutations relative to construct 1 are highlighted. Also shown are the GlcN6P-dependent self-cleavage activities of constructs 1 and 7–12 under standard conditions for 20 h as otherwise described in the legend to . Rate of self-cleavage in the presence of GlcN6P for constructs 1 (circles) and 7 (triangles) under standard conditions

miRNA expression detected by q-PCR and <i>in situ</i> hybridization.

<p>A: Comparison of changes in miRNA expression detected by q-PCR versus microarray analyses for four miRNAs in the SV of C57 and CBA mice. Asterisks indicate statistically significant differences (p<0.05) compared to P21. Each q-PCR plot represents means from three repeats. B: Expression of four miRNAs in the LW of C57 mice using <i>in situ</i> hybridization technique. SV: Stria vascularis. SL: Spiral ligament. 1, 2, and 3 mark the three cell types in the SV (marginal, intermediate, and basal cells), respectively. Bar: 20 µm.</p

Differentially expressed miRNAs in the SV of C57 mice during aging.

<p>The X-axis relates Log2 transformed fold change in miRNA expression at 9 m (in blue) and 16 m (in red) compared to P21. Only those miRNAs which showed a statistically significant different in expression (p<0.05) compared to P21 are shown.</p

EP waveforms and magnitude change during aging.

<p>A. Representative waveforms of EP at three different ages. B. Mean and SD of EP magnitude at different time points during aging. Arrows indicate electrode penetrated into and withdrawn from the scala media.</p

miRNAs that are commonly expressed in the LW of C57 and CBA mice during aging.

<p>Only those miRNAs that showed dynamic changes in the same direction (upregulated or downregulated) in both strains are included.</p

Cross sections of the LW at three different times points during age.

<p>Sections were obtained from cochlear middle turns at P21, 9 m, and 16 m. A. Cross sections stained with Hematoxylin and Eosin. Scale bars: 20 µm. B. Cross sections without any staining. Hyperpigmentation and reduction in the number of capillaries are apparent in the SV at 16 m.</p

Differentially expressed miRNAs in the LW of CBA mice during aging.

<p>The X-axis relates Log2 transformed fold change in miRNA expression at 9 m (in blue) and 16 m (in red) compared to P21. Only those miRNAs whose levels were statistically different (p<0.05) from P21 were included.</p

Network analysis of the relationship between differentially expressed miRNAs and mRNAs in the LW.

<p>A: Upregulated apoptosis-related genes predicted to be targeted by downregulated miRNAs. B: Downregulated apoptosis-related genes predicted to be targeted by upregulated miRNAs. Small circles represent downregulated miRNAs or mRNAs while large circles represent upregulated miRNAs or mRNAs. Purple and orange circles denote differentially expressed miRNAs in C57 and CBA mice, respectively. Red and blue circles represent pro-apoptotic and anti-apoptotic genes, respectively.</p

Transcriptome-wide comparison of the impact of Atoh1 and miR-183 family on pluripotent stem cells and multipotent otic progenitor cells

<div><p>Over 5% of the global population suffers from disabling hearing loss caused by multiple factors including aging, noise exposure, genetic predisposition, or use of ototoxic drugs. Sensorineural hearing loss is often caused by the loss of sensory hair cells (HCs) of the inner ear. A barrier to hearing restoration after HC loss is the limited ability of mammalian auditory HCs to spontaneously regenerate. Understanding the molecular mechanisms orchestrating HC development is expected to facilitate cell replacement therapies. Multiple events are known to be essential for proper HC development including the expression of Atoh1 transcription factor and the miR-183 family. We have developed a series of vectors expressing the miR-183 family and/or Atoh1 that was used to transfect two different developmental cell models: pluripotent mouse embryonic stem cells (mESCs) and immortalized multipotent otic progenitor (iMOP) cells representing an advanced developmental stage. Transcriptome profiling of transfected cells show that the impact of Atoh1 is contextually dependent with more HC-specific effects on iMOP cells. miR-183 family expression in combination with Atoh1 not only appears to fine tune gene expression in favor of HC fate, but is also required for the expression of some HC-specific genes. Overall, the work provides novel insight into the combined role of Atoh1 and the miR-183 family during HC development that may ultimately inform strategies to promote HC regeneration or maintenance.</p></div

Microarray analysis comparing gene expression of mESCs and iMOP cells.

<p>(A) Normalized mean intensity values of differentially expressed genes in untransfected mESC and iMOP cells (n = 3, linear fold change >2 and ANOVA p<0.05). Upregulated genes in iMOP versus mESCs are depicted in red and downregulated genes in green. (B) Hierarchal clustering of gene expression profiles for untransfected mESC and iMOP cells, including profiles for each cell type transfected with indicated expression vector. (C) Number of upregulated genes in transfected mESCs and iMOP cells relative to control pT transfected cells (n = 3, linear fold change ≥1.5 and ANOVA p<0.05). Indicated are upregulated HC-enriched genes (pink) and upregulated non-HC genes (red). (D) Number of downregulated genes in transfected mESCs and iMOP cells relative to control pT transfected cells (n = 3, linear fold change ≥1.5 and ANOVA p<0.05). Indicated are HC-enriched genes (bright green) and non-HC genes (dark green).</p