Figure 2.

The accumulation of TCP17 protein is increased in shade. A, TCP17 transcriptional level is repressed in shade. Seven-day-old Col-0 seedlings grown under Wc were kept in Wc or transferred to shade for the indicated period of time before being harvested for RNA extraction. The error bars represent the SE of three independent biological replicates. *P < 0.05 and **P < 0.01; based on Student’s t test. B, TCP17 protein level is accumulated in shade. Seven-day-old Wc-grown proTCP17::TCP17-GFP transgenic plants were transferred to shade or stayed in Wc for 2 or 4 h. Total protein extracts were separated by SDS-PAGE and analyzed by an immunoblotting approach using an anti-GFP antibody. An immunoblotting assay using antitubulin (TUB) antibody was carried out for a loading control. C and D, TCP17 protein levels were increased, although TCP17 transcription levels were not significantly accumulated, in 35S::TCP17-FLAG transgenic plants after shade treatment. 35S::TCP17-FLAG transgenic plants were treated as shown in B. Total RNA and proteins were extracted for quantitative RT-PCR and immunoblotting assay. Data shown are average of three independent biological replicates and SE. *P < 0.05 and **P < 0.01; NS, not significant (P ≥ 0.05); based on Student’s t test. E, TCP17 is an unstable protein in Wc. Seven-day-old 35S::TCP17-FLAG transgenic seedlings grown in Wc were treated with mock solution, 10 μm CHX, or 10 μm CHX plus 20 μm MG132 for 1 h before the samples were collected for an immunoblotting assay. F, TCP17 is more stable in shade than in Wc. Seven-day-old 35S::TCP17-FLAG transgenic seedlings grown in Wc were transferred to shade for 1 d. The seedlings were kept in shade or exposed to Wc with or without 20 μm MG132 for 4 h. Immunoblotting assays were used to detect protein levels.