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. 2022 May 12;13:2635. doi: 10.1038/s41467-022-30301-y

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© The Author(s) 2022, corrected publication 2022

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Fig. 2 — a Schematic illustrating added components and fluorescently labelled proteins (bold) in experiments shown in b and e. b Representative micrographs of Alexa488-FtsZ (grey) and Cy5-FtsN_cyto (magenta) at increasing FtsA WT and constant FtsZ concentrations. Intensity profiles correspond to the dashed white lines. c Colocalization of FtsZ and FtsN_cyto in the presence of FtsA WT (green) or FtsA R286W (cyan) quantified by PCC. n(WT) = 5/5/3/3 and n(R286W) = 6/4/4/5. d Dynamic colocalization of FtsZ and FtsN_cyto in the presence of FtsA WT (green) or FtsA R286W (cyan) quantified by PCC_diff. n(WT) = 5/5/3/3 and n(R286W);= 4/4/4/3. e Representative micrographs of Alexa488-FtsZ (grey) and Cy5-FtsN_cyto (magenta) at increasing FtsA R286W and constant FtsZ concentration. Line profiles were taken along dashed white lines. f Top: Cy5-FtsN_cyto (magenta) is not enriched on FtsZ filaments after adding Alexa488-FtsZ (grey) and 0.4 µM FtsA WT at 0 min. Bottom: In experiments with 0.4 µM FtsA R286W, Cy5-FtsN_cyto colocalizes well with FtsZ filaments. Right: Increase of PCC after addition of FtsZ and different FtsA WT (top) or R286W (bottom) concentrations. Thick lines depict power law exponential fits towards the mean PCC, shaded area is the standard deviation. g Quantification of the recruitment rate of FtsN_cyto towards FtsZ filaments, extracted from experiments in f by fitting a power law exponential to individual experiments. n(WT) = 3/3/3/3 and n(R286W) = 2/4/3/4. h FtsN_cyto increases the persistency of the FtsZ network with both, FtsA WT and R286W. n(WT/R286W) = 3. i Quantification of the binding affinity of FtsN_cyto towards FtsA WT or R286W by MST. n(WT, R286W) = 3. j Quantification of the duration and frequency of FtsN_cyto single molecule confinement events. n(WT/R286W) = 6/4.Scale bars are 2 µm. Dots in all plots represent independent experiments, thick lines indicate the mean and error bars or shaded areas depict the standard deviation. The boxes indicate the 25–75th percentiles, whiskers show the maximum/minimum values within the standard deviation, the midline indicates the median and diamonds indicate outliers. Source data are provided as a Source Data file.