Figure 4. .

Long RT-PCR and the alternative exons of TAF1. a, Long RT-PCR analysis. The broken line indicates an expected cDNA fragment of TAF1 with the long RT primer on the end of exon 38 (short arrow). By subsequent PCR with the use of the long cDNA, six lanes—TA02, TA08, TA09, TA14, TA15, and TA18 (red bars)—showed multiple bands. Other lanes (black bars) contained single bands. Also shown is the result obtained using the MTS-specific long RT primer on its 3′ end. There was no difference between the TAF1 and MTS primers or between the patient and the control results (patient data not shown). Scale bar=1 kb. b, Ten alternative exons, including two exon skippings and one deletion, were identified by RT-PCR. The detailed sequence information for these exons is annotated in our AB191243 deposition in DNA Databank of Japan (DDBJ). Of 10 alternative exons, 3 were reported elsewhere as exons of MTS, but a form including both exon 32′ and exon 34′ was not detected. For quantitative RT-PCR, 17 TaqMan probes were designed. Two TAF-series probes (red) and 10 TA-series probes (pink) were designed to detect mainly the TAF1 common forms and alternative splicing isoforms, respectively. The other five MTS-series probes (orange) were designed to detect the MTS transcripts reported elsewhere. Var. = variant. Scale bar=5 kb.