Fig. S5.
DDB2−AGO1 complex. (A) Root growth assay. Seven-day-old WT (Col), ddb2-3, ddb2-3/DDB2-FLAG, and ddb2-3/DDB2K314E-FLAG expressing plants were exposed to 900 J/m2 of UV-C; t test *P < 0.01; **P < 0.05. (B) In vivo pull-down of AGO1 with DDB2-FLAG protein upon UV-C exposure; ddb2-3/DDB2-FLAG and ddb2-3/DDB2K314E-FLAG expressing plants were used for IP assays using anti-FLAG antibody. WT (Col) plants were used as negative control. Coomassie blue staining of the blot is shown. (C) In vivo pull-down of AGO1 with DDB2-FLAG protein upon UV-C exposure; ddb2-2/DDB2-FLAG and atr ddb2-2/DDB2-FLAG expressing plants were used for IP assays using anti-FLAG antibody. WT (No) plants were used as negative control. Coomassie blue staining of the blot is shown. (D) Immunoblot analysis of DDB2 and AGO1 protein contents upon UV-C exposure in chromatin extracts from WT and atr plants. Anti-histone H3 and anti-UGPase antibodies were used as controls for insoluble (P, chromatin) and soluble (S) fractions, respectively. Signal intensity relative to H3 is indicated below each lane. (E) Same as D for AGO1 protein in ddb2 plants. (F) In vivo pull-down of AGO1 with DDB2 protein upon UV-C exposure in chromatin fraction; ddb2-2/DDB2-FLAG plants were used for IP assays using anti-FLAG antibody. WT plants were used as negative control. Anti-histone H3 and anti-UGPase antibodies were used as controls for insoluble (P, chromatin) and soluble (S) fractions, respectively. Coomassie blue staining of the blot is shown. (G) Tandem ChIP (Tandem-ChIP) of AGO1 and DDB2-FLAG, upon UV-C exposure, at two hot spots in ddb2-3/DDB2-FLAG expressing plants using anti-AGO1 followed by anti-FLAG antibody. As negative control for ChIP, WT plants were used as well as actin2 region. Data are presented as enrichment (±SD) of the IP signal and are representative of two independent biological replicates; t test *P < 0.01 compared with time point 0.