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. 2016 Feb 8;170(4):2365–2377. doi: 10.1104/pp.15.01283

Figure 6.

Figure 6.

OsMADS23, OsMADS27a, and OsMADS57 bind to the CArG motifs of the OsRDR1 promoter. A, Schematic structures of the effector and reporter constructs for the Y1H assay. OsMADS23, OsMADS27a, and OsMADS57 were fused to the GAD, and the reporter gene LacZ was driven by the OsRDR1 promoter fragment containing a CArG motif. B, OsMADS23, OsMADS27a, and OsMADS57 bound to the promoter fragment of OsRDR1 in yeast. C to E, EMSAs showed that OsMADS23, OsMADS27a, and OsMADS57 bound to the CArG motif of the OsRDR1 promoter. The biotinylated probe containing the CArG motif sequence was incubated with GST-OsMADS23, GST-OsMADS27a, or GST-OsMADS57, while the probe incubated with no protein or GST protein was used as a negative control. Nonlabeled probes were used as cold competitors. F and G, ChIP-PCR assays showed that OsMADS23, OsMADS27a, and OsMADS57 bound to the OsRDR1 promoter regions containing CArG motifs (red boxes) in rice plants. Two pairs of PCR primers (P1/P2 and P3/P4) are indicated, and the amplified PCR bands are indicated by arrows. Asterisks indicate the dimers formed by primers. Ab, Antibody.