Fig. 6: NONO facilitates the splicing mature and stabilizes hypoxia-induced genes.
A Number of transcripts bound by NONO under normoxia and hypoxia in HepG2. B Distribution of NONO RIP-sequencing peak annotation for different regions. C, D Gene enrichment of biological process (C) and enriched KEGG pathways (D) analysis for the NONO-bound mRNA. E Significantly enriched RNA motifs among the RIP-seq. F Lysate from HepG2 cells after 1% O2 or Cocl2 (100 μM) treatment for 24 h was subjected to RIP assay, then RIP assay products were extracted by trizol. The mRNA bound by NONO were quantified by RT-qPCR with the indicated pairs of primers. G HepG2 cells under hypoxia were treated with actinomycin D (2 μM) and harvested at the indicated time points, then RNA was extracted from these cells and quantified by RT-qPCR with the indicated pairs of primers. H, I Lysate from HepG2 cells after 1% O2 or Cocl2 (100 μM) treatment for 24 h was subjected to RIP assay, then RIP assay products were extracted by trizol. The snRNA U6 (H) and pre-mRNA (I) bound by NONO were quantified by RT-qPCR with the indicated pairs of primers. J HepG2 cells with NONO knockout or not were treated with hypoxia (1% O2, 24 h) or normoxia. The expression ratio of Cas9 and NONO-KO group in intron and mRNA of indicated genes were determined by RT-qPCR.
