Fig 8. ClnR directly and specifically binds several DNA targets.
Electrophoretic mobility shift assays were performed as described in Methods using His-tagged ClnR and fluorescein-labeled DNA encompassing regions upstream of A) clnR, B) vanZ, C) CD1606, or D) sigU. ClnR was added to reactions at varying concentrations (specified in nM in A, in μM elsewhere) either without or with 0.5 μM LL-37, as indicated. Competitive EMSAs were performed with the addition of unlabeled target DNA (specific) or unlabeled Pspo0A DNA (non-specific) at either 10x or 100x the concentration of labeled target DNA. 125 nM ClnR was used for the competitive EMSA for Pcln, 8 μM for all others. Apparent Kd values were calculated as described in Methods. Graphs are the binding curves showing the mean and standard deviation from three independent replicates.