PPP3CA isoforms were identified by aligning genomic sequence with human PPP3CA ESTs using Sequencher software (Version 3.0, Gene Code Co.). As described previously (Liu et al., 2006), exon-specific primers and fluorescent FAM-labeled MGB probes were designed (Table S3) across two alternatively spliced exons using Primer Express (Applied Biosystems) to avoid nonspecific amplification of genomic DNA. The cDNA templates were amplified using ABI Prism 7900HT Sequence Detection System (Applied Biosystems) with a cycling program of 50°C for 2 min, 95°C for 10 min, followed by 40 cycles of 95°C for 0.25 min and 60°C for 1 min. Predeveloped human glyceraldehyde-3-phosphate dehydrogenase (GAPD) (Applied Biosystems) provided a control. Quantification used methods in user bulletin #2 (ABI Prism 7700 Sequence Detection System) and graphs used Prism 3.0 (GraphPad Software, Inc. San Diego, CA).
