Figure 2

DNA and RNA expression of introduced bc. (a) DNA and RNA were isolated from a wild-type βc-transfected clone 5C and four βc79-transfected clones mE, m821, m951, and 63m. The isolated DNA and reverse-transcribed RNA were subjected to the amplification of βc and GAPDH by PCR. Primers 1S/1A, 2S/2A, and 3S/2A were used for detection of βc. cDNA obtained from 5C was diluted by 10 times, while cDNA from βc79-expressing clones were directly applied to PCR without dilution. M: Marker. (b) The βc-specific transcripts were quantified by real-time PCR using primer 1S/1A. The amounts of βc-specific transcripts relative to those of endogenous GAPDH transcripts were compared among three wild-type and four βc79 clones. The average values from 3 runs of real-time RT-PCR are presented. Error bars express S.E.M. (c) The mE and 5C cells were cultured in the presence of 25 μg/mL DRB and the ratios of βc-specific transcripts to GAPDH transcripts were plotted at various time points. *: significantly lower than the ratio from 5C cells at the same time points (p < 0.05).