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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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