2.9 TriFC Assay
In the case of TriFC assay, the full-length coding sequences for MeCSP5
and MSCP were cloned into pSAT4-nEYFP-C1 and pSAT4-cEYFP-C1 vectors,
respectively. The sequence of 6xMS2 was fused with CRIR1 or
control lncRNA under the control of the 35S CaMV promoter in the
pCAMBIA1301 vector. For the TriFC assay in rice protoplasts, transient
transfection of rice protoplast cultures were performed according to the
protocol described previously (Li, Xiang, Yang, Chen & Zhang, 2015).
Fluorescence was detected using confocal laser scanning microscope
(Olympus FluoView FV1100) 16-20 h after transfection.