Fig. 4: SOX9 deletion modifies alternative splicing of Srsf5.

A Transcript level analyses of reads from the RNA-seq data showing the abundance of transcripts of coding (blue box plots) and non-coding isoforms (orange box plots) in controls and Ins-Cre;Sox9-/- islets. The exon compositions of the splice isoforms are shown. RNA-seq was carried out on islets isolated from control or Ins-Cre;Sox9-/- animals (13-16 months old, n = 4 per group). Data are presented as box plots (center line at the median, upper bound at 75th percentile, lower bound at 25th percentile) with whiskers at minimum and maximum values. Each dot represents one animal. B Splicing event in Srsf5 that leads to inclusion of a poison cassette exon with a premature termination codon. C Sashimi plots of alternative splicing in Srsf5. RNA-seq was carried out on islets isolated from control or Ins-Cre;Sox9-/- animals (13-16 months old, n = 4 per group). Data are presented as box plots (center line at the median, upper bound at 75th percentile, lower bound at 25th percentile) with whiskers at minimum and maximum values. Each dot represents one animal. D PCR validation of the comparison of abundance of transcripts with increased levels of intron 5 inclusion in Srsf5 between control and knockout islets. Primer locations for each product are shown. Controls, n = 3 from two independent cohorts, Knockouts, n = 3 from three independent cohorts. E Quantification of the semi-quantitative PCR shown in (D). RNA was isolated from control and knockout animals (n = 3 per group). Error bars represent S.E.M. using a two-tailed Student’s t-test, *p < 0.05, **p < 0.005, ***p < 0.0005. F Proximity Ligation Assay showing a close association between Sox9 and Y14 in Ins1 cells was carried out two independent times. Size bar, 50 µm. Source data are provided as a Source Data file.