Real-time PCR was performed using an Eppendorf Realplex2 machine and Sybr-Green (BioRad), with the default parameters (95°C 2 min; 95°C 15 sec, 55°C 15 sec, 68°C 20 sec; repeat 40x) except that the 68°C elongation was lengthened to 40 sec. Maximal rates of amplification per cycle were calculated for all primer pairs and all samples in all assays and averaged 1.98 ± 0.06 (SD) for a set of 8 consecutive 96-well assays. Data are calculated with respect to GAPDH for each sample within an assay; since values with respect to GAPDH for each transcript were consistent across assays, data were averaged. Use of actin instead of GAPDH for normalization did not alter the results. Primers are listed in Table 1.
