The numbers of human heart samples used for qRT-PCR for each genotype were given under each column.
We thank Elvira Arza for confocal microscopy, Toni Diez and Vanessa Blanco for help with histology and for heart samples, Aida Garcia and Piedad Fernandez for help with lentivirus production and Laura Carramolino for embryo dissection, and Simon Bartlett for editorial work.
Heart samples were incubated for 90 min. Incubation was stopped by rapid vacuum filtration over Whatman GF/C glass microfibre filters (VWR International SAS, Strasbourg, France) presoaked with polyethylenimine 0.3% in order to reduce non-specific binding to the filters.
Five to ten μg of RNA from heart samples was used for miRNA microarray.
The same heart samples were divided for 2-dimensional polyacrylamide gel electrophoresis analysis or Western blot analysis.
Plasma and reconstituted heart samples were analyzed for NE content using a modified inline capture, column-switching high performance liquid chromatography procedure with electrochemical detection [ 23, 27].
Expression was normalized to eye tissue (fish) or heart (due to lack of eye samples for female frog and chicken).
DNA was extracted using the Maxwell 16 system (Promega Corporation, Madison, WI) for cerebrum, spleen, liver, pancreas, kidney, lung, bone marrow, jejunum, or using the DNeasy kit (Qiagen, Valencia, CA) for heart samples.
To evaluate the expression level of the myomesin genes in pathological situations in general, heart samples of mouse models for HCM or DCM were examined by RT-PCR analysis.
In general, brain samples showed high TSS activity for the Dp140 and Dp71 + Dp40 isoform groups (Fig. 4), while the muscle and heart samples showed high TSS activity for the Dp427 isoforms group.
