README for Figure 3B+S5A.csv 
*** This file contains the raw data obtained on DNA and DNA-Rok Δ75-96 complexes using tethered particle motion represented
in Figure 3B+S5A 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 3: The linker domain of Rok is important for its cooperative DNA binding. A) Sequence alignment of 
Rok wildtype (NCBI accession number: NP_389307.1), Rok Δ75-96 and sRok (NCBI accession number: YP_004243533.1). 
The N- and C-terminal domain are indicated with a black box and the linker domain with a red line. B) RMS values 
obtained as a function of Rok Δ75-96 concentration as measured by TPM in the presence of 50 mM KCl. The RMS values
were determined from fitting with a Gaussian distribution. For reference, Rok WT data is shown (reproduced from 
figure 2B).  Error bars represent the propagated standard deviation from at least two independent measurements. 
C) DNA occupancy (fraction of Rok-bound tethers of the total amount of DNA tethers) as function of protein 
concentration in nM. The data points were fitted using the Hill binding model.

Legend Figure S5: Rok Δ75-96 compacts and bridges DNA but with decreased cooperativity. A)  Histograms of Root 
Mean Square displacement (RMS) obtained for 32%GC DNA as a function of Rok Δ75-96 at concentrations of 0, 1, 2, 3, 
4, 5, 7, 8, 9, 10, 11, 12, 13, 14 and 15 nM as measured by TPM in the presence of 50 mM KCl. The histograms were 
fitted to Gaussian distributions, in which the RMS value at ~150 nm represents bare DNA and the population with 
an RMS at ~100 nm represents DNA bound by Rok Δ75-96. The bare DNA and Rok-DNA complex populations are highlighted 
with a light blue and magenta box, respectively. The data for each concentration originates from at least two 
independent measurements. B) DNA recovery (%) as a function of protein concentration in µM in the presence of 50 
mM KCl at 25°C. For reference, Rok WT is shown (reproduced from figure 2A). Data are plotted as mean values and 
the error bars represent the standard deviation from three independent measurements. Dashed lines are lines to 
guide the eye. 

 *** The data were obtained using Tethered particle motion as described in the associated article. 

*** Data obtained for each protein concentration(s) is given in this table as plotted in figure 3B+S5A.
Column A: Rok Δ75-96(nM) = Concentration of Rok Δ75-96 in nanomolar
Column B: Replicate = number of replicate
Column C: RMS (nm) = Root mean square deviation in nanometer measured for each bead 
Column D: rho = anisotropic ratio (movement in x-direction/movement in y-direction) 
Column E: stdev = standard deviation 
Column F: FoV = field of view (data with the same FoV were collected in one image) 
Column G: bead = beads in each FoV are represented by a unique number 
The values given in column D and E are tether quality criteria and used for data filtering as described in the associated article.