Figure 2

Detection of transcripts of Tgfbr1 variants by RNase protection analysis. Total RNA extracted from CA1109 effusion tumor cells were protected by corresponding antisense riboprobes, as indicated at the top. The riboprobes were synthesized by T7 RNA polymerase using T7 promoter-containing DNA fragment of individual variant as a template. The predicted protected fragments are 263 bp for D1 (DM1), 192 bp for D2 (DM2), 151 bp for D3 (DM3), and 90 bp for D4 (DM4). Lanes 5 and 6: the mixed riboprobes (M) were incubated with (+) or without (-) RNase A/T1, respectively. Lanes 1–4: the total RNA (10 μg aliquots) was hybridized with individual purified antisense riboprobes and the protected fragments were resolved on a 6% acrylamide/8 M urea sequencing gel. * Indicates an unidentified Tgfbr1 transcript variant in CA1109 tumor cells protected by the anti-DM3 riboprobe.