Cardiac Gene Expression Profiling

Real-time PCR was performed as previously described [11 (link)]. Total RNA was extracted from cardiac tissue (peri-infarction area, ventricular septum, and remote area) and reverse transcribed using TaqMan reverse transcription reagents (Applied Biosystems, Stockholm, Sweden). RT-PCR was performed on an ABI PRISM 7700 system (Applied Biosystems) using rat-specific primers for VEGF-A, transforming growth factor-β (TGFβ), interleukin (IL)-1β, and IL-6 (Assay ID: Rn01511601_m1, Rn00572010_m1, Rn00580432_m1, and Rn01410330_m1, respectively) in the first examination; for SDF1 (Assay ID: Rn00573260_m1) in the second examination; and for GFP (Assay ID: Mr04097229_mr) in the third examination. Each cDNA sample was evaluated in duplicate. Expression of target genes was normalized to that of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for each sample. Relative gene expression was determined using the 2^−ΔΔC_T method.