Fig. 7

dhx38 modulates the alternative splicing of a subset of genes involved in mitosis and DNA damage. (A) Semi-qPCR confirms the abnormal splicing of the genes shown in Fig. 6D. The left band represents gene splicing in the wild-type siblings, whereas the right band represents splicing in the dhx38 mutants. Percent spliced in, PSI; skipped exon, SE; retained intron, RI; exon, E; intron, I; premature termination codon, PTC. Black underlines denote abnormally spliced exons/introns. Experiments were performed with three replicates; gapdh was used as the internal control. (B) qRT-PCR analysis confirming mRNA expression of differentially spliced genes shown in Fig. 6D. n≥12 per group, performed with three replicates; gapdh was used as the internal control. Data show the mean±s.d. Significance was determined using two-tailed unpaired Student's t-test; *P<0.05; **P<0.01; ***P<0.001. (C) The genes shown here are those with splicing abnormalities and downregulated expression, which were by semi-qPCR and qRT-PCR, respectively. Of the genes assessed, cenpk, smc5, knl1, kdm8, eme1, tonsl, ccng2, ccnb2 and aaas are predicted to undergo PTC-NMD events, whereas trip12, scrib, pard3ab, sept6, ift20 and lzts2a undergo differential isoform transition, with mis18a, dtl, rad9a, mak and cep131 generating disordered isoforms. (D) Western blotting for CCNB2 in the si-DHX38-641 displays decreased protein expression compared with the si-NC or si-DHX38-325 group. n=3, *P=0.041.