2.7 ChIP analysis
ChIP experiments were performed as described by Buzas et al. (2011). The
non-crosslinked leaf sheath of the CRCT-FLAG overexpression line was
used for this experiment. FLAG-CRCT associated DNA was
immunoprecipitated using anti-DYKDDDDK tag antibody beads (Fujifilm Wako
Chemical). Immunoprecipitated DNAs and input DNAs were purified using
QIAquick PCR Purification Kit (Qiagen). ChIP-qPCR was performed using
gene-specific primers (Table S1). Purified immunoprecipitated DNAs and
input DNAs were used as templates, and qPCR was carried out using
TBgreen Premix Ex Tag GC (Takara) and a thermal cycler (MyGoPro, IT-IS
Life Science Ltd.) according to the manufacturer’s instructions. The 18S
ribosomal RNA gene (XR_003238822) was examined as a reference.