Figure 5.

YY2 K247 methylation regulates YY2 binding with chromatin. (a, b) Inducible HeLa cells expressing pRevTRE-Flag-HA-YY2 were treated with doxycycline (Dox, 600 μgml⁻¹) for 48 h, followed by ChIP-seq to detect YY2-binding sites in the genome. Peak finding (a) and motif analysis (b) were performed using HOMER. (c) YY2 binding detected by ChIP-seq was shown for ABL1, TP53/p53 and RAD1 genes as indicated. (d) HeLa cells were transfected with vectors expressing HA-tagged YY2 (wt) or YY2 (K247R), followed by ChIP with anti-HA antibody and qPCR with primers specifically targeting promoter regions of selected genes as indicated. ChIP signals were presented as fold induction over wt after being normalized to input (±s.e.m., ***P<0.001). (e) Control (wt), SET7/9 or LSD1 KO HeLa cells were transfected with vector expressing HA-tagged YY2, followed by ChIP with anti-HA antibody and qPCR with primer specifically targeting promoter regions of selected genes as indicated. ChIP signals were presented as fold induction over wt after being normalized to input (±s.e.m., *P<0.05, **P<0.01, ***P<0.001, NS, nonsignificant). (f) HeLa cells were transfected with vectors expressing HA-tagged YY2 in the presence or absence of SET7/9 (wt), SET7/9 (m), LSD1 (wt) or LSD1 (m), followed by ChIP with anti-HA antibody and qPCR with primer specifically targeting promoter regions of selected genes as indicated. ChIP signals were presented as fold induction over CTL (YY2 alone) after being normalized to input (±s.e.m., *P<0.05, **P<0.01, ***P<0.001).