Fig. 3.

MORC2 recruits DNMT3A to promote DNA methylation at the NF2 and KIBRA loci. a DNMT3A is required for MORC2 to suppress NF2 and KIBRA mRNA levels. HepG2 cells were infected with control or MORC2-expressing (Lv-MORC2) lentivirus, together with scramble siRNA or siRNAs targeting DNMT1 (si-DNMT1), DNMT3A (si-DNMT3A), or DNMT3b (si-DNMT3b) for 48 h. The relative levels of NF2 and KIBRA mRNA were determined by qRT-PCR (n = 3 per biological repeats). b Western blot analysis of anti-MORC2 and anti-DNMT3A immunoprecipitate demonstrates interaction between MORC2 and DNMT3A. c MORC2 is required for DNMT3A to interact with NF2 and KIBRA promoters. The occupancy of DNMT3A at the promoter regions of NF2 and KIBRA loci was determined by ChIP-qPCR. d–e MeDIP-qPCR analysis of DNA methylation status of the NF2 and KIBRA loci in control or MORC2-expressing HepG2 cells transfected scramble siRNA or si-DNMT3A (d), or in MORC2^+/+ or MORC2^−/− PLC cells infected with control or DNMT3A-expressing lentivirus (e). f DNMT3A is required for MORC2 to suppress expression of NF2 and KIBRA. Western blot analysis of NF2 and KIBRA in control or MORC2-expressing HepG2 cells, or cells that were further transfected with scramble siRNA or siRNAs targeting DNMT1, DNMT3A, and DNMT3B. Normal IgG was included as negative control for ChIP-qPCR and MeDIP-qPCR. β-actin was used as a loading control. Data are shown as the mean ± SD (a, c, d, e) of triplicates and representative images (b, f) are presented. *P < 0.05