Figure 4.

Nuclear runoff analysis. Nuclei were isolated from A673 and SJRH5, and nuclear transcription was performed with [α-³²P]UTP. ³²P-labeled RNA was purified and hybridized to linearized plasmids immobilized onto slot blots. (A) PAX3 intron 7/exon 8 sequences used as 5′- and 3′-PAX3 hybridization probes. Exons 6–8 are represented by the black boxes. The 5′-and 3′-PAX3 hybridization probes are indicated by the brackets below the horizontal line. The t(2;13) breakpoint region in SJRH5 is indicated by the arrow. (B) ³²P-labeled RNA from A673 lacking the t(2;13) (Left) and SJRH5 containing the t(2;13) (Right) was hybridized to filters containing either 5′-or 3′-PAX3 hybridization probes. Plasmids GAPDH and pBluescript served as internal and negative controls, respectively. (C) Relative transcription levels were determined by normalizing hybridization signals for the number of uridines in the sense strands of the 5′- or 3′-PAX3 hybridization probes and calculating a 5′- or 3′-PAX3 to GAPDH ratio. Averages and standard deviations were calculated from three experiments.