Fig. 4
NSUN2 methylates HIP1 RNA and regulates its expression. (A) IGV tracks from m5C-meRIP-seq analysis showed m5C enrichment of HIP1. (B) Expression of HIP1 in patients with MM and healthy controls from GSE80608. (C) The correlation analysis of mRNA expression between NSUN2 and HIP1. (D) HIP1 mRNA expression in patients with MM and healthy controls detected by RT-qPCR. ***p < 0.001. (E) NSUN2 knockdown decreases the level of HIP1 mRNA in RPMI8226 cells. **p < 0.01. (F) Abundance of methylated HIP1 detected by RNA m5C RIP-qPCR from mock and shNSUN2 RPMI8226 cells. m5C RIP-qPCR with IgG antibody experiments are performed as negative controls. (G) Half-life of HIP1 mRNA measured after actinomycin D treatment. ***p < 0.001, **p < 0.01, *p < 0.5. NSUN2, NOP2/Sun RNA methyltransferase family member 2; IGV, Integrative Genomics Viewer; HIP1, huntingtin interacting protein 1; RT-PCR, reverse transcription PCR; m5C RIP-qPCR, m5C methylated RNA immunoprecipitation quantitative PCR.
