Figure 4.

EIN3 Protein Is Stabilized by Cold Stress.

(A) Cold treatment stabilizes the EIN3-GFP protein. 35S:EIN3-GFP seedlings (7 d old) in the ein3 eil1 mutant background (EIN3-OE/dm) were treated at 4°C for 0, 3, 6, and 12 h before the tissues were used for detecting GFP fluorescence. Treatment with ACC (10 μM) for 6 h was used as a positive control. Bars = 50 μm.

(B) Immunoblot analysis of EIN3-GFP under cold treatment. EIN3-OE/dm seedlings (7 d old) were treated with ACC (10 μM) or cold for 6 h, and the proteins were extracted and subjected to immunoblot assays using an anti-GFP antibody.

(C) Immunoblot analysis of EIN3-GFP under cold stress in the presence of AVG and Ag⁺. EIN3-OE/dm seedlings (7 d old) grown on MS medium with AVG (2 μM) or AgNO₃ (20 μM) were treated at 4°C for 0, 3, 6, and 12 h.

(D) Immunoblot analysis of EIN3-GFP protein in 7-d-old 35S:EIN3-GFP seedlings in the ein3 eil1 and ein2 ein3 eil1 backgrounds after cold stress. Rubisco, ribulose-1,5-bisphosphate carboxylase/oxygenase.

(E) Immunoblot analysis of EBF1-GFP protein in 7-d-old 35S:EBF1-GFP seedlings after cold treatment.

(F) Immunoblot analysis of EBF1-TAP protein (detected by anti-Myc antibody) in 7-d-old 35S:EBF1-TAP ein2 seedlings after cold treatment.