5–2.0% isoflurane (Minrad Inc.) in an air:O2 (4:1) mixture. The MRI experiments Selleckchem Akt inhibitor were performed on a 9.4 T small animal MRI system (Bruker BioSpin MRI) equipped with a gradient system capable of 400 mT/m using established procedures [26]. For MR signal transmission and reception, a circular polarized birdcage resonator with an inner diameter of 21 mm was used. Scout images of the heart anatomy were acquired for accurate planning of the subsequent cine cardiac scans. For left ventricular function analysis, high-resolution black-blood short-axis images covering the entire left ventricle

were acquired using the self-gating technique IntraGate (Bruker BioSpin MRI), which is based on a fast low-angle shot (FLASH) multislice sequence with an extra navigator echo [51]. The following XL184 molecular weight parameters were used for data acquisition: field-of-view (FOV) = 25 × 25 mm2, matrix dimension = 128 × 128 (zero-filled to 256 × 256), spatial resolution = 98 × 98 μm2, six to seven contiguous slices of 1.0 mm thickness, pulse angle = 10°, echo/repetition time (TE/TR) = 1.8/50.5 ms, number of repetitions (NR) = 200, total acquisition time = 21.5 min. In post processing, acquired MR image data was

assigned to 10 cardiac phases and the end-expiratory motion state according to the self-gating signal. MRI images were analyzed to determine end diastolic volume (EDV) and end systolic volume (ESV) using Paravision 5.0 (Bruker BioSpin) and subsequently stroke volumes (SV) and EFs were calculated from the obtained values. All statistical analyses were performed with Prism 4.0 (Graphpad Software Inc.). Data were analyzed with the nonpaired Student’s t-test assuming that the values followed a Gaussian distribution. A p value of 17-DMAG (Alvespimycin) HCl < 0.05 was considered as significant. We would like to thank Eva Allgäuer for technical support. This study received financial support from the Swiss National Science Foundation (116499 and 130823 to B.L.) and from the Austrian Genome Research Programme GEN-AU II and III (Austromouse) to T.R. The authors declare no financial or commercial conflict of interest. Disclaimer:

Supplementary materials have been peer-reviewed but not copyedited. Figure S1. Isolation and characterization of a myhca614-629-specific TCR. (A) Myhca peptide-stimulated effector T cells were fused to BW 5147 lymphoma cells. Proliferating clones were subcloned by limiting dilution and expression of CD4 and particular Vβ chains was assessed by flow cytometry. Representative dot plots of clone 35 with monoclonal subclones 5.4 and 5.4 are shown. (B) Antigen-specificity of subclones 5.4 and 5.5 was confirmed by IFN-γ ELISPOT assay using dendritic cells pulsed with myhca614-629 or unpulsed (med.) as stimulators. (C) Schematic illustration of the DNA sequence of the myhca-specific TCR that was obtained following PCR sequencing and database alignment. The sequence of the CDR3 region is depicted in detail. Figure S2. Lack of cardiac myosin alpha expression in thymi of BALB/c mice.