FIGURE 3.

Flowering Locus C is stabilized by HPY2 in vivo. Double transgenic plants containing 35S-FLC-FLAG₃ and XVE-HA₃-HPY2 (A) or 35S-mFLC (K154R)-FLAG₃ and XVE-HA₃-AtSIZ1 (B) were incubated in liquid medium with β-estradiol to induce HPY2 expression. After incubation for 15 h, HA₃-HPY2, FLC-FLAG₃, and mFLC-FLAG₃ levels were assessed by Western blotting with anti-HA or anti-FLAG antibodies. Tubulin was used as a loading control. Numbers under lanes indicate relative intensities. Protein levels were normalized to a value of 1.00 for FLC or mFLC levels without inducer (“–” in both panels). RNA concentrations of FLC-FLAG₃ and mFLC-FLAG₃ were determined by real-time qRT-PCR using a FLAG primer and a gene-specific primer. Tubulin RNA was used as a loading control. (C) Graphical expression of Tubulin, FLC-FLAG₃, and mFLC-FLAG₃ transcript levels from (B). Bars indicate standard errors (n = 3).