MSE of the different projection and the original projection.

<p>MSE of the different projection and the original projection.</p

Inhibition of G3bp1 restricts ET-1 induced cardiac hypertrophy.

<p>(a) Neonatal myocytes were infected with adenoviruses expressing miR-SC (control virus), G3BP1, miR-1 or 10% FBS, as indicated. Total protein lysate was extracted and analyzed by western blotting for Eif4e, rasGAP, Actin and G3bp1 (n = 2). (b) Neonatal myocytes cultured were treated with adenovirus expressing miR-1, G3BP1 or control virus, as indicated. After 24hrs the cells were supplemented with [H³]-leucine to the medium. After additional 24hrs, protein and DNA was extracted and counts per minute (CPM) measured, normalized to DNA, averaged and plotted. Graph represents relative CPM/μg of DNA Error Bars represents SEM, * represents p<0.05 vs. control. # represents p<0.05 G3bp1+miR-1 vs. miR-1 (n = 3). (c) Immunocytochemistry was performed on myocytes plated in glass chamber slides and treated with Ad-LacZ or Ad-G3BP1, as indicated. Myocytes were stimulated with ET-1 in the presence or absence of Ad-siG3bp1 or Ad-siLUC. The cells were fixed and stained for phalloidin and DAPI. N = 3. d. The graph represents cell area quantified using Image J, error bars represents SEM and * is p<0.05 vs. Ad-siLUC, and # is p<0.05 vs. ET-1+Ad-siLUC, n = 3 independent experiments.</p

The flowchart of the iterative FBP algorithm.

<p>The flowchart of the iterative FBP algorithm.</p

Parallel Projection: an object <i>f</i>(<i>x</i>, <i>y</i>) and its projection <i>p</i>_<i>θ</i>(<i>t</i>) from angle <i>θ</i>.

<p>Parallel Projection: an object <i>f</i>(<i>x</i>, <i>y</i>) and its projection <i>p</i>_<i>θ</i>(<i>t</i>) from angle <i>θ</i>.</p

MiR-1 expression is posttranscriptionally regulated during cardiac hypertrophy.

<p>Pool of three sham/adult and TAC operated mouse hearts were used for anti-RNA pol II, anti-H3K9ac and anti-Gtf2b ChIP-Seq. Binary analysis files (BAR) from the ChIP-Seq data was viewed in Affymetrix’s Integrated Genome Browser (IGB), which shows the fragment densities of RNA pol II, H3K9ac and Gtf2b (<i>y-</i>axis) aligned in 32–50 nucleotide bins along the chromosomal coordinates (<i>x</i>-axis) for miR-1-133 clusters. The arrow indicates the transcription start site and the direction of transcription. (a) IGB images of miR-1-1 and miR-133a-2 transcript with RNA polII, H3K3ac and Gtf2b densities across intergenic regions of chromosome 2. (b) IGB images of miR-1-2 and miR-133a-1 cluster with RNA polII, H3K3ac and Gtf2b densities within the Mindbomb 1 (Mib1) gene located in chromosome 18. (c) Total mRNA extracted from mouse hearts from sham or TAC operated hearts for 1day or four days were used for qPCR analysis of primary, pre-miR-1-1, pre-miR-1-2 and mature miR-1 levels. The results were normalized to 18S (primary and pre- transcript) or U6 (mature) and shown as ratio of TAC/sham. Error bars represents standard error of mean (SEM) and * is p< 0.05, n = 3. (d) Neonatal myocytes cultured in growth-inhibited (serum free) conditions were stimulated with 100nM endothelin-1 or 10%FBS for 1hr or 24hrs, as indicated. Total RNA extracted was used for qPCR analysis of primary, pre- and mature miR-1. Error bars represents SEM, and * is p<0.05 and ** is p< 0.005. N = 3.</p

Eif4e is direct target of miR-1 in cardiomyocytes.

<p>(a) Targetscan and Pictar predicted miRs and sites on Eif4e 3’UTR that are broadly conserved between vertebrates. (b) Neonatal myocytes cultured in growth-inhibited medium (serum-free, SF) were infected with adenovirus constructs of Eif4e, as indicated, with multiplicity of infection (MOI) of 5 or 20 for 24hrs before protein extraction, followed by Western blotting for Eif4e and Gapdh. Relative Eif4e represents densitometry of two independent blots, averaged and normalized to Gapdh. (c) The graph represents relative increase in Eif4e protein expression levels with Eif4e-Mut and Eif4e-Cds constructs compared to Eif4e-Fl. Error bars represents SEM and ** is p<0.001, n = 3. (d) Neonatal myocytes were infected with Ad-SC, Ad-MiR-1 or Ad-anti-miR-1 in increasing doses in the presence or absence of 10% FBS, respectively, as indicated. Western blot analysis was performed on protein lysate for Eif4e and MHC. n = 3. (e) The graph represents change in Eif4e protein expression with Ad-miR1- or Ad-anti-miR-1 normalized to MHC. Error bars represents SEM and * is p<0.05, n = 3. (f) Immunocytochemistry was performed in myocytes plated in glass chamber slides and stimulated with ET-1 for 24hrs in the absence or presence of exogenous miR-1. Cells were stained for Eif4e, MHC and DAPI. Eif4e signal was quantified using Analyze particles tool from Image J. The graph represents average number of Eif4e stained particles/cell and average size of particles with the treatments. Error bars represent SEM, and * is p<0.05 and ** is p<0.005. Three independent experiments, four field each with more than 2cells used for quantification.</p

G3bp1 regulates miR-1 processing in cardiomyocytes.

<p>(a) Neonatal myocytes were treated with Ad-LacZ or Ad-G3bp1 for 24hrs; total RNA extracted was used for qPCR analysis for rno-primary and rno-pre-miR-1. The results were normalized to 18S, averaged and plotted. Error bars represents SEM, and * is p<0.05, n = 3. (b) Neonatal myocytes cultured in serum free conditions were treated with Ad-LacZ, Ad-G3bp1, Ad-anti-miR-1 (two doses, as indicated) or 10% FBS for 24hrs before extracting total RNA for Northern Blots. Blots were probed for miR-1, anti-miR-1 sequence or U6. 5S is shown as loading control. (c) Myocytes were cultured and treated with adenovirus expressing shRNA against G3bp1 (Ad-siG3bp), Ad-siLUC (control virus) or 10% FBS, as indicated. Northern blots were performed and probed for miR-1 and miR-21 (specificity control), 5S is shown as loading control. (d) Northern blots from 3a and b, and two independent blots were scanned, quantified, averaged and plotted. The error bars represents SEM and * p<0.05 vs. SF control. (e) Western blot analysis of the cytoplasmic and nuclear fractions of protein lysate was performed on myocytes for the indicated genes on myocytes treated with Ad-lacZ (control virus), Ad-G3bp1 or 10% FBS, as indicated for 24hrs. (f) Western blot analysis of cytoplasmic and nuclear fractions of protein lysate was performed on myocytes treated with Ad-siLUC (control virus), Ad-siG3BP1 or 10% FBS, as indicated. (g) Total protein lysate from cultured myocytes treated with Ad-lacZ (control virus), Ad-G3bp1 or Ad-miR-1, as indicated were separated by SDS-PAGE and protein expression of selected genes analyzed, n = 2. (h) Western blots were quantified, averaged and plotted. Error bars represents SEM and * p<0.05 vs. respective control. All experiments were performed in triplicates, unless indicated otherwise and representative blots have been shown.</p

The compare of reconstruction performance between SIRT, SART, MAP-EM and Iterative FBP (the image size is 512 × 512 and the angle interval is 0.5°).

<p>The compare of reconstruction performance between SIRT, SART, MAP-EM and Iterative FBP (the image size is 512 × 512 and the angle interval is 0.5°).</p

A Novel Iterative CT Reconstruction Approach Based on FBP Algorithm - Fig 7

<p>(a) The compare of MSE using SART and Iterative FBP. (b) is the portion of the first 60s of (a).</p

Reconstruction from the filtered projection: the linear interpolation procedure.

<p>Reconstruction from the filtered projection: the linear interpolation procedure.</p