Fig. 6

Reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis of splicing status shows differences between transcripts. a Diagram showing the location of diagnostic amplicons. Exons are denoted by blue boxes and the intron is represented by a black line. Amplicons are indicated by lines below. Pre-mRNAs were detected using oligonucleotides that amplify the exon–intron boundary at the 5′ splice site. b Relative pre-mRNA levels of three transcripts, RPL28, RPL39 and RPS13, analyzed by RT-qPCR, normalized relative to steady state (SS) levels. Data show how the level of each amplicon approaches the level detected at steady state as labeling time increases. Data were normalized to the levels of exon 2 and steady state to account for different RNA yields obtained at each labeling time. Different transcripts show different rates of splicing. c-e UCSC genome browser screen shots showing the change in distribution of reads at different labeling times (y-axis) for RPL28, RPL39 and RPS13, with annotation below in blue. Exons are represented by blue boxes and intron indicated by a blue line. SS indicates steady-state levels, generated by sequencing total RNA