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Fig. 3 | Genome Biology

Fig. 3

From: Regulation of constitutive and alternative mRNA splicing across the human transcriptome by PRPF8 is determined by 5′ splice site strength

Fig. 3

Altered splicing of transcripts encoding proteins required for mitotic progression after PRPF8 depletion. a RT-PCR analysis of RNA from PRPF8-depleted cells reveals a variety of splicing defects in transcripts that encode critical factors required for mitotic progression, including retained introns (CDC20 and Separase), and alternative terminal exons (APC8). RT-PCR analysis of CDC20, Separase, and APC8 using the primers indicated in the schematic are shown alongside the corresponding coverage plots (control siRNA in black, PRPF8 siRNA in red). The splicing defects are indicated by arrowheads. b, c Quantitative RT-PCR analysis of RNA from PRPF8-depleted cells, including retained introns (CDC20, Separase, and NUDC), skipped exons (ASPM and SKA2), and alternative terminal exons (APC8). Plots are relative to RNA levels in control siRNA-treated cells, assigned an arbitrary value of 1, and show the mean of triplicate readings from three independent experiments ± standard error of the mean. d Protein expression analysis of genes with altered splicing following PRPF8 depletion

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