Ricle obtained in WT/3M, Myo-Tg and Myo-3M mice. Final results are presented because the imply SEM and CD66e/CEACAM5 Proteins Gene ID represent four different mice (p 0.001 compared with the Myo-Tg mice).J Mol Biol. Author manuscript; offered in PMC 2009 September 5.Young et al.PageNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptFigure 2. NF-B activation cascades Myo-3M mice hearts(A) Nuclear protein was extracted from the hearts of WT/3M, Myo-Tg mice and Myo-3M. Binding reactions had been performed with an NF-B oligonucleotide labeled with 32P-dATP. The complex formation was eliminated with excess unlabeled NF-B oligonucleotide. The complex formation was confirmed by supershift evaluation working with p65 antibody. NE: Nuclear extract. (B) Quantification of EMSA utilizing an arbitrary density unit (10 /NE). (C) Western blots profile of NF-B p65 protein inside the nucleus. Histone antibody was utilized as an internal nuclear protein loading manage. (D) Expression of p65 Muscle-Specific Kinase (MuSK) Proteins Accession active protein in the heart section of each Myo-Tg and Myo-3M mice and were photographed with an Olympus photomicroscope at 20 magnification. This figure is representative of 3 different mice in each group (WT/3M andJ Mol Biol. Author manuscript; available in PMC 2009 September 5.Young et al.PageMyo-Tg). (E). Cytoplasmic protein extracts were produced from both WT, 3M, Myo-Tg and Myo-3M mouse hearts at 24 weeks of age. Tissue extracts (50 ) had been analyzed for the intracellular degree of total IB protein content material and (F) Actin protein was applied as an internal loading control. Outcomes are presented because the imply SEM and represent 3 various mice in each group (Myo-Tg and Myo-3M (p 0.001).NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptJ Mol Biol. Author manuscript; offered in PMC 2009 September five.Young et al.PageNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptFigure 3. Determination of steady state degree of ANF, -MHC and MLC2 (v) gene expressions in 3M miceTotal RNA was extracted from hearts of 24-week old WT/3M, Myo-Tg and Myo-3M mice. mRNA expression was determined working with (A) ANF, (B) -MHC, (C) MLC2 (v) and (D) 18S rRNA oligonucleotides labeled with 32P-ATP as a probes. Benefits are presented because the mean SEM and represent three various mice (p 0.001 compared with the Myo-Tg mice).J Mol Biol. Author manuscript; accessible in PMC 2009 September 5.Young et al.PageNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptJ Mol Biol. Author manuscript; accessible in PMC 2009 September 5.Figure four. Determination of steady state level of TNF, IL-1 and IL-6 in Myo-3M mice heartsTotal RNA was extracted from hearts of 24-week old WT/3M, Myo-Tg and Myo-3M mice. mRNA expression was determined using (A) TNF, (B) IL-1 and (C) IL-6 oligonucleotide labeled with 32P-dATP as a probe. (D) 18S rRNA probe was employed as a loading control. Outcomes are presented as the imply SEM and represent three unique mice (p 0.001 compared with all the Myo-Tg mice).Young et al.PageNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptFigure 5. Analysis of macrophage infiltration in Myo-3M mice heartsTotal RNA was extracted from hearts of 24-week old WT/3M, Myo-Tg and Myo-3M mice. Semi-quantitative RT-PCR was performed employing (A), F4/80 (B) MCP-1 and (C) MCAF particular primers. Results are presented as the imply SEM and represent three various mice (p 0.001 compared with the Myo-Tg mice). (D). Immunohistological analysis of MCP-1 in cardiac section of WT/3M, M.
