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. 2019 Aug 7;116(34):17105–17114. doi: 10.1073/pnas.1907968116

Fig. 4.

Fig. 4.

CsBRC1 directly binds to the auxin efflux carrier gene CsPIN3 to repress its expression. (A) Schematic diagram of the putative cis-elements in CsPIN1b (P1) and CsPIN3 (P3A–P3E) used for CsBRC1 binding assay. (B) Yeast 1-hybrid assay showed binding of CsBRC1 to P1, P3A, P3B, P3C, and P3E. The numbers in the right corner indicate the basal concentration of Aureobasidin A (AbA; ng/mL). (C) Luciferase activity measurement in tobacco leaves after coexpression of Pro35S:CsBRC1 and ProCsPIN1b:LUC or ProCsPIN3:LUC. The empty vector 62-SK was used as the control. Four independent transfection experiments were performed. (D) Callus images from cucumber hypocotyl explants transfected with empty control, Pro35S:GFP, and Pro35S:CsBRC1-GFP used in ChIP assay. (E) ChIP-qPCR assay showed that CsBRC1 binds to P3A, P3B, and P3C regions of CsPIN3. Three technical replicates and 3 biological replicates from different calli were performed. Error bars represent ±SD. (F) EMSA assay showed that CsBRC1 binds to P3A, P3B, and P3C regions of CsPIN3. Significance analysis was conducted with the 2-tailed Student’s t tests (**P < 0.01).