Measurement of catalase activity in vector control and CDF4RNAi plants. Ten‐day‐old green seedlings (“Young”) and the third and fourth rosette leaves from 36‐day‐old plants (“Old”) were used, respectively. Three independent experiments were conducted. Data are represented as means ± SD, n = 3. Student's t‐test, *P < 0.05.
qPCR analysis of CAT genes expression in the third and fourth rosette leaves from 36‐day‐old plants. The expression of CAT genes in the wild‐type plant is given as 1. Three independent experiments were conducted. Data are represented as means ± SD, n = 3. Student's t‐test, *P < 0.05.
Schematic diagram indicating the locations of the putative CDF4‐binding motif clusters (D1–D5) in the promoter of CAT2.
ChIP‐qPCR analysis of the relative binding of CDF4 to the promoter of CAT2. An anti‐HA monoclonal antibody was used for DNA immunoprecipitation from 32‐day‐old pER8::CDF4‐HA transgenic plants. Black columns indicate the enrichment fold changes normalized to that of ACT2. Three independent experiments were conducted. Data are represented as means ± SD, n = 3. Student's t‐test, *P < 0.05.
Transient dual‐luciferase reporter assay. The pGreenII‐0800 LUC construct containing the CAT2 promoter and the p62‐SK construct with or without CDF4 were transiently co‐transformed into Col‐0 protoplasts. Firefly luciferase (LUC) and Renilla luciferase (REN) activity were measured after culturing the protoplasts under low light conditions for 16 h. Six independent experiments were conducted. Values are given as mean ± SD, n = 6. *P < 0.05 by Student's t‐test.
