Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA, USA
Arrhythmia and sudden death associated with elevated cardiac chloride channel activity
Version of Record online: 24 OCT 2011
© 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd
Journal of Cellular and Molecular Medicine
Volume 15, Issue 11, pages 2307–2316, November 2011
How to Cite
Ye, L., Zhu, W., Backx, P. H., Cortez, M. A., Wu, J., Chow, Y.-H., Mckerlie, C., Wang, A., Tsui, L.-C., Gross, G.J. and Hu, J. (2011), Arrhythmia and sudden death associated with elevated cardiac chloride channel activity. Journal of Cellular and Molecular Medicine, 15: 2307–2316. doi: 10.1111/j.1582-4934.2010.01243.x
- Issue online: 24 OCT 2011
- Version of Record online: 24 OCT 2011
- Accepted manuscript online: 14 DEC 2010 07:38AM EST
- Received: May 28, 2010; Accepted: December 8, 2010
Fig. S1 Southern Blot analysis of CFTR transgenic mice. (A) Identification of transgenic mouse lines. (B) Estimation of transgene copy numbers (1.5 to 20 copies) based on the amount of genomic DNA and size of the mouse genome relative the plasmid control used in the analysis. The probe used was K18 intron 1 shown in Fig. 1A. Abbreviations: WT, wild-type; PC, positive control (plasmid DNA).
Fig. S2 Analysis of transgenic expression of the human CFTR in mouse heart tissue. (A) Expression of the human CFTR detected by in situ hybridization with an antisense probe described in 'Methods'. (B) A serial section of the same heart hybridized with the sense control RNA. (C) A heart section from a nontransgenic littermate probed with the same antisense probe shown in (A). (D), (E) and (F) Enlarged areas from (A), (B) and (C).
Fig. S3 Levels of human (h)CFTR transgene expression in heart tissues of male and female CFTR transgenic mice determined by TaqMan realtime RT-PCR. Total RNA (1 μg) was reverse transcribed using random hexamers and SuperScript2 reverse tran scriptase (Invitrogen) following the manufacturer's protocol. Twenty nanograms of the resulting cDNA were used in real-time PCR performed with a PCR machine from ABI (Prism 7700, Applied Biosystems, Foster City, CA, USA). The sequences of the primers and probe used in the real time PCR are as following: forward primer, CCGAATTCCCGCGTCA; reverse primer, GGAGACAACGCTGGCCTTT; and probe, CAAATACTTCCACCATGGAGAGGTCGCC. A dilution series was used to determine the efficiency of amplification of each primer/probe set, allowing the relative quantification method to be employed (Livak and Schmittgen, Methods 25, 402--408, 2001). 18S was used as a reference gene to normalize the hCFTR expression. Data are expressed as values of 2--ΔCt × 103versus 18S and are means ± S.E.M. from six mice in each group. There is no human CFTR transgene expression in transgenic negative littermates.
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