Skip to main content
. 2016 Dec 5;173(1):155–166. doi: 10.1104/pp.16.01024

Figure 5.

Figure 5.

SUF4 binds to all five EC1 promoters. A, Gel-shift assay without (lane 1) and with 10 (lane 2), 50 (lane 3), 100 (lane 4), 200 (lane 5), and 400 ng (lane 6) of recombinant 6xHIS-SUF4-STREPII added to a radioactively labeled 108-bp EC1.1 promoter fragment covering the DNA region used as bait in the yeast one-hybrid screen. B, Gel-shift assay with 50-fold (50×) and 100-fold (100×) excess of unlabeled EC1.1 promoter fragment as a cold competitor added to the reaction mix with 200 ng of 6xHIS-SUF4-STREPII. The control reaction is without cold competitor (0×). C, Fifty and 150 ng of recombinant 6xHIS-MBP-SUF4, and 150 ng of 6xHIS-MBP as a control, were mixed with 10 ng of radioactively labeled EC1 promoter fragments. Lane 1, Radioactively labeled promoter fragment only; lane 2, radioactively labeled promoter fragment with 150 ng of 6xHIS-MBP tag only; lane 3, radioactively labeled promoter fragment with 50 ng of 6xHIS-MBP-SUF4; lane 4, radioactively labeled promoter fragment with 150 ng of MBP-SUF4; lane 5, radioactively labeled promoter fragment with 150 ng of MBP-SUF4 and 100-fold excess of cold competitor (unlabeled promoter fragment). Asterisks mark free probes, arrows mark shifted bands of protein-DNA complexes.