Expression Analysis of VIN3 Gene Family Members during the Course of Vernalization.
(A) mRNA expression analysis of VIN3 gene family members during the course of vernalization using real-time RT-PCR. NV, nonvernalized; 10V, 10 d of vernalization; 20V, 20 d of vernalization; 30V, 30 d of vernalization; 40V, 40 d of vernalization; 40VT10, 40 d of vernalization followed by 10 d of normal growth temperature. Relative fold change was determined by normalization with the levels of PP2A gene. Real-time PCR experiments were performed with three biological replicates, and error bars indicate sd. Primer sequence information used for real-time RT-PCR analysis is shown in Supplemental Table 1 online.
(B) Immunoblot analysis of VIN3 family of proteins during the course of vernalization. NV, nonvernalized; 40V, 40 d of vernalization; 40VT10, 40 d of vernalization followed by 10 d of normal growth temperature. Corresponding insertional mutants were used as negative controls for VIN3, VIL1/VRN5, and VIL2/VEL1 immunoblots. Two independent RNAi lines (#5 and #9) were used as negative controls for VIL3/VEL2 immunoblots. WT, the wild type.