Journal: Theranostics

Article Title: Remote postconditioning ameliorates stroke damage by preventing let-7a and miR-143 up-regulation

doi: 10.7150/thno.48135

Figure Lengend Snippet: Heat map of microRNA expression profiles in the whole ischemic area from sham-operated, tMCAO and tMCAO + RLIP animals. (A) The average of signal intensity values for each significantly expressed miRNA is reported (p < 0.05). Red and green indicate up- and down-regulation respectively, according to the colorimetric scale in the right side of the panel. a, b and c indicate the three experimental group: (a) Sham, (b) tMCAO and (c) tMCAO + RLIP animals. Each vertical line refers to three samples for each experimental group. (B) Volcano plot analysis in the lower side of panel shows the comparison of miRNA levels after tMCAO and RLIP induction. The y-axis corresponds to the mean expression value of log 10 (p-value), and the x-axis displays the log 2 fold change value. The blue dots represent the miRNA levels expressed in rat brain 24 h after stroke induction; the orange dots represent the miRNA levels expressed in rat brain 24 h after tMCAO + RLIP.

Article Snippet: RNA samples were sent to LC Sciences (Houston, Texas, USA), a global biotechnology company that is provided with a microRNA microarray service.

Techniques: Expressing, Comparison

Journal: Theranostics

Article Title: Remote postconditioning ameliorates stroke damage by preventing let-7a and miR-143 up-regulation

doi: 10.7150/thno.48135

Figure Lengend Snippet: Validation of microarray results by real-time PCR. MicroRNA levels analysed by Real-Time PCR in ischemic brain regions from rats subjected to tMCAO and tMCAO + RLIP are expressed as fold change over the respective sham-operated controls. Each column represents the mean ± S.E.M. Results of microRNAs expression were normalized with respect to 4.5S RNA as internal control. n = 3 or 4 per group. S is for Sham-operated group.

Article Snippet: RNA samples were sent to LC Sciences (Houston, Texas, USA), a global biotechnology company that is provided with a microRNA microarray service.

Techniques: Biomarker Discovery, Microarray, Real-time Polymerase Chain Reaction, Expressing, Control

Journal: PLoS ONE

Article Title: Microarray Analysis of microRNA Expression during Axolotl Limb Regeneration

doi: 10.1371/journal.pone.0041804

Figure Lengend Snippet: Quantitative PCR validates miR-21 microarray results. Two first columns compare the averaged fold change between 17 dpa blastema (Bl) and stump (St) samples for LNA based qPCR assays (yellow bar), and for previous microarray data (red bar). Also, the individual fold changes between Bl and St for the three animals (biological replicates) are shown (blue bars). The relative miR-21 expression was calculated based on the efficiency corrected ΔΔCt method and normalized with miR-20a and miR-200b . The y-axis is a log scale. A fold change >1 indicates upregulation in Bl compared to St samples [ p ≤0.03 (qPCR), p ≤0.0001 (array); two-tailed t- test]. Inset , illustrates how non-isotopic Northern blot using digoxigenin-labeled LNAs against hsa-miR21 and hsa-U6 (control) are useful to detect the axolotl versions of these small RNAs. The axolotl miR-21 ( Amex-miR-21 ) is detected as a band of ∼20 nt which is over expressed in blastema when compared with stump and blood samples.

Article Snippet: After running the RNA samples on a gel and confirming their integrity, 1.2 μg were sent to LC Sciences (Houston, TX) to be processed using their MicroRNA detection Microarray Service, and to be hybridized to a custom vertebrates chip (MRA-2001).

Techniques: Real-time Polymerase Chain Reaction, Microarray, Expressing, Two Tailed Test, Non-Isotopic Labeling, Northern Blot, Labeling, Control

Journal: PLoS ONE

Article Title: Microarray Analysis of microRNA Expression during Axolotl Limb Regeneration

doi: 10.1371/journal.pone.0041804

Figure Lengend Snippet: A. Comparison between the human and axolotl miR-21 target sites in their Jagged1 genes. Nucleotide alignment of the miR-21 target site-containing region present in the 3′-UTR of the human Jagged1 ( Hsa-Jag1 , NM_000214) and the axolotl Jagged1 ( Amx-Jag1 , JF907581). The 22 bases comprising the target site for miR-21 are in yellow except for the 7 bases complementary to the miR-21 seed region (green color). Vertical bars (|) denote nucleotide identities between the two sequences. The numbers at both sides of the alignment are the nucleotide position on each 3′-UTR. B. In vitro luciferase assays testing the effect of miR-21 on the 3′-UTR of Amex-Jag1 in axolotl cells . Results are expressed as percent of co-reporter-normalized Renilla activity against reference vectors. Bars denote standard error of mean of the amount of independent assays. Student t- test was done and the obtained p- values determined that the Target of Hsa-miR-21 is a good positive control as biosensor for the activity of this microRNA ( p ≤0.005; two-tailed t- test). These results suggest that Amex-Jag1 may in fact be a target for miR-21 because significant differences were found in the Renilla signal recorded from cells electroporated with only the vector containing the 3′-UTR of Amex-Jag1 , versus the cells that were also electroporated with Pre-miR-21 *** (p≤0.005) but not Pre-miR30a. When the latter experiment was repeated with the mutant, seedless - Amex-Jag1- 3′-UTR, any sensitivity to exogenous miR-21 was lost.

Article Snippet: After running the RNA samples on a gel and confirming their integrity, 1.2 μg were sent to LC Sciences (Houston, TX) to be processed using their MicroRNA detection Microarray Service, and to be hybridized to a custom vertebrates chip (MRA-2001).

Techniques: Comparison, In Vitro, Luciferase, Activity Assay, Positive Control, Two Tailed Test, Plasmid Preparation, Mutagenesis