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. 2000 Jul;157(1):59–68. doi: 10.1016/s0002-9440(10)64517-8

Figure 1.

Figure 1.

Nonexpression of the PrP gene in a skin fibroblast cell line established from the PrP−/− mice. Two cell lines were established from primary cultures of skin fibroblasts isolated from the PrP−/− mice (SFK) or from the PrP+/+ mice (SFH). They were maintained for 4 months in vitro before starting the experiments. The cells were incubated for 72 hours in LS medium with (SFH-B and SFK-B) or without (SFH-C and SFK-C) inclusion of 50 ng/ml of bFGF for the last 24 hours before processing for RNA preparation. Fifty nanograms of cDNA prepared from total RNA by reverse transcription (RT) was amplified by PCR using primer pairs specific for the PrP gene (lanes 1−8) or the β-actin gene (lanes 9−16) listed in Table 1 ▶ . Lanes 1 and 9 represent SFH-C cells processed for RT-PCR omitting the RT step; lanes 2 and 10, SFH-C cells processed for RT-PCR including the RT step; lanes 3 and 11, SFK-C cells processed for RT-PCR omitting the RT step; lanes 4 and 12, SFK-C cells processed for RT-PCR including the RT step; lanes 5 and 13, SFH-B cells processed for RT-PCR omitting the RT step; lanes 6 and 14, SFH-B cells processed for RT-PCR including the RT step; lanes 7 and 15, SFK-B cells processed for RT-PCR omitting the RT step; lanes 8 and 16, SFK-B cells processed for RT-PCR including the RT step. The DNA size marker (bp) is shown on the left.