Fig. 1.

Clinical relevance of PKM splicing and it’s correlation with BORIS. (a) Schematic representation of PKM spliced isoform, the cancer-specific PKM2 isoform containing exon 9 whereas normal PKM1 isoform contains exon 10 as has been represented in the processed mRNA with the primer sets directed against the specific exon as a whole, exon junction-specific primer sets were used to specifically measure the spliced isoforms. The exon 9 inclusion was indicated by the exon junction primers exon 8–9/9, whereas exon 10–11/11 indicates the inclusion of exon 10. (b) RPS16 normalized qRT-PCR in paired normal and tumor HNC patients samples using the indicated exon junction specific primers for PKM gene (n = 10). (d) MeDIP in paired normal and tumor HNC patients samples and qRT-PCR of exon 10 region in PKM gene relative to input (n = 4). (e-f) ChIP analysis in paired normal and tumor tissues of HNC patients using (c) RNA Pol II and (d) BORIS antibody, followed by qRT-PCR relative to input (n = 3). Graphs show mean values ± SD. P as calculated using two-tailed Student’s t-test,*P < 0.05, ** P < 0.01, *** P < 0.001, ns = non-significant