real-time quantification of target gene and of the housekeeping gene was
performed in separate reactions. The threshold cycle (Ct) values
obtained by real-time RT-PCR were used to calculate the accumulation
of target gene (relative mRNA accumulation) relative to
α-tubulin transcript by the 2^–ΔΔCt method, where ΔΔCt = (Ct
target gene – Ct α-tubulin). Results were based on the
average obtained for at least two replicate RT-PCR reactions per
sample.