Journal: The Journal of Biological Chemistry
Article Title: Critical Role of Nuclear Calcium/Calmodulin-dependent Protein Kinase II?B in Cardiomyocyte Survival in Cardiomyopathy *
Figure Lengend Snippet: Reduction of CaMKIIδB transcripts and protein after Dox treatment in isolated cardiomyocytes. A , expression levels of CaMKIIδB and Dox-sensitive mRNAs in primary neonatal rat cardiomyocytes were analyzed by quantitative RT-PCR. B , rapid and severe reductions of CaMKIIδB transcripts in cardiomyocytes treated with Dox are shown. Expression of the indicated mRNAs was measured after the indicated time points of Dox exposure by real time PCR (IQ SYBR green, Bio-Rad). The data are corrected for GAPDH expression level and normalized so that expression at 0 h equals 1. Results shown are representative of three independent experiments. C ). The results are representative of three separate experiments. D , specificity of the band migrating at 55 kDa was demonstrated by transfecting primary cardiomyocytes with siCtr or siCaMKIIδB, and Western blot analysis was performed 72 h later using the CaMKIIδ-specific antibody. Untr , untreated; cTnI , cardiac troponin I.
Article Snippet: Real time PCRs were performed after RT-PCR (RETROscriptTM , Ambion) with various dilutions of cDNA template using IQ SYBR Green and an Opticon 2 reader (Bio-Rad).
Techniques: Isolation, Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, SYBR Green Assay, Western Blot