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. 2017 Feb 24;13(2):e1006642. doi: 10.1371/journal.pgen.1006642

Fig 8. RID1 directly bind to the promoter regions of Hd3a and RFT1.

Fig 8

(A) Gel shift assays of His and His-RID1 recombinant proteins interacting with promoter region of Hd3a and RFT1. Escherichia coli–produced recombinant RID1 protein were incubated with biotin-labeled Hd3a and RFT1 in the absence or presence of 100- or 500-fold molar excess of the unlabeled probes as competitor for the electrophoretic mobility shift assay (EMSA) reaction and analyzed by electrophoresis. The fragment with mutated core cis-element served as the negative control. (B) ChIP analysis of transgenic plants expressing RID1-FLAG-HA fusion protein. Nuclei from RID1-FLAG-HA transgenic plants’ leaves were immune precipitated by anti-HA. The precipitated chromatin fragments were analyzed by qPCR using four primer sets amplifying Hd3a and RFT1 regions (I, II, III, and IV), as indicated in Fig 7A. The input (without antibody precipitation) chromatin was analyzed and used as the control. The ChIP experiments were repeated two times using independent biological replicates with similar results, and one representative data set is shown.