Fig. 4

Confirmatory RNA sequencing in P1 and P3. For both A and B, in the top panel, paired end reads from the RNA sequencing BAM file are displayed for the proband. In the lower panels, the RNA sequencing read pileup tract is displayed with the novel (orange) and known (blue) junctions annotated in the proband and in aggregated data from GTEx controls, respectively. Beneath, the gene transcript isoforms are displayed. A, RNA sequencing analysis performed on blood in P1 compared to normalized GTEx blood samples (n = 755) (21). The results for ASNS (displaying exon 9 and 10) demonstrate evidence of splice disruption due to a deep intronic indel (indicated by the red box in the proband) with cryptic exon creation and intron 9 read-through. B, RNA sequencing analysis performed on an EBV-transformed lymphoblastoid cell line (LCL) in P3 compared to normalized GTEx lymphocyte samples (n = 174). The results for TCF4 (displaying exon 10 to 13) demonstrate evidence of splice disruption due to a near-splice variant (indicated by the red line in the proband) with skipping of exon 11 in approximately 20% of reads. E, exon