README for Figure 4F.csv 
*** This file contains the raw data obtained on DNA, DNA-Rok complexes using bridging assay experiments represented in Figure 4F of 
Article: The B. subtilis Rok protein is an atypical H-NS-like protein irresponsive to physico-chemical cues
Authors: Erkelens, Qin, van Erp, Miguel-Arribas, Abia, Keek, Markus, Cajili, Schwab, Meijer and Dame
Journal:  
DOI:
Corresponding author: rtdame@chem.leidenuniv.nl; wmeijer@cbm.csic.es
 

Legend Figure 4:The bridging activity of sRok can be modulated by salt concentration A) RMS values obtained for 
32%GC DNA as a function of sRok concentration as measured by TPM in the presence of 50 mM KCl.  The RMS values 
were determined from fitting with a Gaussian distribution. Error bars represent the propagated standard deviation 
from at least two independent measurements.  Some error bars are hidden behind the data points. B) DNA recovery 
(%) as function of (s)Rok concentration in the presence of 50 mM KCl at 25C. For reference, Rok WT is shown 
(reproduced from figure 2A). Data are plotted as mean values from three independent measurements and the error 
bars represent the standard deviation. Dashed lines are lines to guide the eye. C) Structural predictions using 
Alphafold of  Rok homodimer, sRok homodimer and Rok:sRok heterodimer (left to right). The protein structures are 
colored by the predicted lDDT-Ca (PLDDT) values with the following color scheme: blue (> 90), light blue (90 to 
70), yellow (70 to 50), orange (< 50). The PLDDT indicates the local confidence in the predicted structures from 
0-100, with 100 corresponding to highest confidence. D) SDS-PAGE analysis of His-tag pull down assay. Rok 6xhis 
was captured on HisPur Ni-NTA Magnetic beads and, when applicable, sRok was added in 1:1 molar ratio. E) DNA 
recovery (%) measured in the presence of 50 mM KCl + 1 mM MgCl2 at 25C with different ratios Rok:sRok. The total 
amount of protein used was constant at 0.5 M. F) DNA recovery (%) measured in the presence of 1 mM MgCl2 at 25C 
and 0.5 M protein with different KCl concentrations. 

*** The data were obtained using bridging assay experiments as described in the associated article. 

***The data labeled overview represents the values plotted in the graph of figure 2C. The raw data contains the individual replicates.
Column A: Sample number
Column B: Presence of bait DNA in sample
Column C: Presence of 32P-labeled prey DNA in sample
Column D: Amount of KCl in incubation buffer in millimolar
Column E: Protein added (0.5 uM)
Column F: Counts per minutes 
Column G: DNA recovery (%)

Columns B, C, D and E indicate the contents of the sample. Column F indicates the radioactivity of the sample. 
Column G indicates the DNA recovery, which is calculated by subtracting the value in column F of the control 
(sample minus bait DNA) from the sample itself. This value is then divided by the values in column F of sample 1 
and multiplied by 100%. 









