Analysis of TCF1 expression and nuclear localization of the TCF1 protein.

(A) Three-week-old Col-0 (WT) plants were subjected to low temperature (4°C) and the samples were harvested at the indicated time points for semi-quantitative RT-PCR analysis of TCF1 transcripts. ACT2 (At3g18780) was used as a loading control. (B) GUS staining of transgenic plants expressing TCF1pro::GUS under normal temperature or treated at 4°C for 7 days. (C) Semi-quantitative RT-PCR for TCF1 in different tissues with or without cold treatments for 7 days. R, root; Sh, shoot; L, leaves; F, flowers; Si, siliques. (D) Semi-quantitative RT-PCR analysis for TCF1 of three-week-old Col-0 plants treated with 100 μM ABA, 400 mM mannitol, 20% PEG6000, 300 mM NaCl for 3 h and 4°C Cold treatment for 24 h. (E) Localization of fluorescence in tcf1-1 plants expressing a TCF1pro::GFP-TCF1 fusion at 22°C (Upper) and 4°C for 7 days (Bottom). GFP: GFP-TCF1 fusion protein (tcf1-1TCF1-3), DAPI: DAPI staining, Merge: Merger of GFP and DAPI channels (Scale bars, 20 μm).