Panel A: Epicardial transplantation of myoblast sheets (Sheet group, n = 18) significantly improved the left ventricular ejection fraction (EF).

<p>In sham-treated animals (Control group, n = 19), EF decreased markedly during the 2-week follow-up. This decrease in EF was attenuated by myoblast intramyocardial injection therapy (Injection group, n = 17). Panel B: Significant improvement in fractional shortening (FS) was observed in the Sheet group. FS significantly deteriorated in the Control group, while in the Injection group this decrease was attenuated. Panel C: As a measure of left ventricular remodeling, the left ventricular end-diastolic diameter (LVEDD) decreased in the Sheet group and increased in the Control group. In the Injection group, the LVEDD remained similar to the baseline value. Panel D: Representative hematoxylin-eosin-stained paraffin-embedded sections of the heart on the midventricular short axis from each group. The myocardium in the Sheet group demonstrated less remodeling of the left ventricle than the Control group. A similar, but less pronounced, effect on remodeling was evident in the Injection group. * p < 0.05, ** p < 0.01 Sheet group vs. Control group; † p < 0.05, †† p < 0.01 Sheet group vs. Injection group.</p

Panels A-C: Representative immunofluorescence images of CD11b-expression, with DAPI counterstaining of the nuclei of the myocardium 2 weeks after myoblast cell therapy (4 weeks after myocardial infarction).

<p>Infiltration of CD11b-positive cells by epicardial transplantation (panel A) was similar to that of the Control group (panel C). Panel D: Clusters of CD11b-positive inflammatory cells can be seen in the myocardium of the Injection group at a higher magnification. Panel E shows densitometry quantitation of CD11b-expression in the various groups (n = 6). In the group receiving intramyocardial injections, an increased leukocyte infiltrate was evident in comparison to the Control or Sheet groups (*** p < 0.001 vs. Injection group). Panels F-I show CD68-staining similar to that of CD11b. Quantitation of the CD68-positive area showed increased CD68-positive cell infiltration in the Injection group in comparison to the Sheet and Control groups (panel J, * p < 0.05 for both comparisons).</p

Electropotential mapping was performed epicardially with a 16-needle probe at 2.0-ms intervals.

<p>In the premedication maps, one cardiac R-R-cycle is shown (images from 18 ms to 218 ms) at a normal rat heart rate (300 beats per minute, bpm). An area of reentry can be seen as red coloration in the middle of the border area measured. Sixty seconds after a bolus injection of isoproterenol, the reentry area was observed, while 120 s after the isoproterenol bolus, a full ventricular tachycardia has developed, with a heart rate of approximately 6000 bpm.</p

Ventricular premature contractions (VPCs) after epicardial myoblast sheet transplantation (Sheet group, n = 18) or intramyocardial myoblast injections (Injection group, n = 17).

<p>The Control group (n = 19) received, instead of myoblast therapy, a sham operation 2 weeks after myocardial infarction. Electrocardiography was monitored continuously. The number of VPCs was significantly higher in the Injection group on days 1 (p < 0.05) and 14 (p < 0.01) after myoblast transplantation than in the Control group (*) and significantly higher on days 7 (p < 0.05) and 14 (p < 0.01) than in the Sheet group (†). There was no ventricular tachycardia recorded in any of the animals.</p

Study protocol timeline.

<p>Myoblast cell therapy as intramyocardial injections or epicardial sheets was administered 2 weeks after induction of heart failure and ischemia by ligation of the left anterior coronary artery (Myocardial infarction). Electrocardiography (Holter) was monitored constantly, using implanted Holter monitors, and specific recordings were made on days 1, 7, and 14 after cell therapy administration. Echocardiography (Echo), as measured first before and then 2 weeks after myoblast transplantation, was used to evaluate therapy efficacy. Epicardial electropotential mapping (EEPM) was performed 14 days after therapy administration.</p

Myocardial gene expressions of inflammatory genes in the sham-treated (Control group, n = 6) and in the groups receiving myoblast cell therapy as intramyocardial injections (Injection group, n = 6) or epicardial cell sheets (Sheet group, n = 6).

<p>Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01 compared with the Control group. † p < 0.05 compared with the Injection group.</p

Additional file 8: of Intracellular signalling pathways and cytoskeletal functions converge on the psoriasis candidate gene CCHCR1 expressed at P-bodies and centrosomes

Supplementary Information and Figure S2. Information about qPCR and co-localization of CCHCR1 with P-body markers. Lists of pre-designed TaqMan Gene Expression Assays and nucleotide sequences of self-designed qPCR primers. Counting the colocalization of CCHCR1 with P-body markers in the CCHCR1-HEK293 cell lines and calculation of p-values for the comparison between cell lines. Figure S2. Îł-tubulin staining of the CCHCR1 cells. Antibody against Îł-tubulin was used as a marker for centrosomes. (PDF 1046 kb

Additional file 1: of Intracellular signalling pathways and cytoskeletal functions converge on the psoriasis candidate gene CCHCR1 expressed at P-bodies and centrosomes

Table S1. Microarray data and gene enrichment analysis of the CCHCR1 Iso1 cell lines. Gene expression profiling data of microarrays from the Isoform 1 CCHCR1-HEK293 cell lines. Gene enrichment analysis of differentially expressed genes using the KEGG pathway analysis of WebGestaltR and functional cluster analysis of the DAVID. (XLSX 305 kb

Additional file 4: of Intracellular signalling pathways and cytoskeletal functions converge on the psoriasis candidate gene CCHCR1 expressed at P-bodies and centrosomes

Table S4. Isoform- and haplotype-specific gene enrichment with the shared DEGs of the CCHCR1-HEK293 cell lines. Gene enrichment analyses of DEGs shared by only the Non-risk (Diff N), Risk (Diff R), isoform 1 (Diff iso1), or isoform 3 (Diff iso3) CCHCR1cell lines (see in detail Fig. 4 Venn diagram). The DEGs shared by all the CCHCR1 cell lines (Intersection) were analyzed as well. Analyses were done using the GO and cluster analyses from DAVID and KEGG pathway analysis from WebGestalt and WebGestaltR. (XLSX 307 kb

Additional file 2: of Intracellular signalling pathways and cytoskeletal functions converge on the psoriasis candidate gene CCHCR1 expressed at P-bodies and centrosomes

Table S2. Differentially expressed genes of RNAseq from the comparisons between the CCHCR1-HEK293 cell lines and controls. Up- and downregulated DEGs of CCHCR1-overexpressing cell lines Iso1Non-risk (1 N), Iso1Risk (1R), Iso3Non-risk (3 N), and Iso3Risk (3R) when compared with controls (wild type and vector transfected cells). (XLSX 1215 kb