Journal: eLife

Article Title: A conserved cell division protein directly regulates FtsZ dynamics in filamentous and unicellular actinobacteria

doi: 10.7554/eLife.63387

Figure Lengend Snippet: ( A and B ) Still images from and showing the localization of FtsZ-YPet in sporulating ( A ) wild-type (WT; SS12) and ( B ) Δ sepH mutant hyphae (MB750). Arrow heads in ( B ) point at aberrant spores or gaps in FtsZ-YPet-ladders (filled arrow head) or indicate lysed hyphae (open arrow heads in DIC image). Note that DIC images correspond to a later time point than fluorescence micrographs to show the terminal sporulation phenotype. Scale bars: 10 µm. ( C ) and ( D ) Kymograph analysis of FtsZ-YPet dynamics during sporulation-specific cell division in WT ( C ) and ΔsepH hyphae ( D ), ectopically expressing an additional copy of ftsZ-ypet from the native promoter (SS12 and MB750). The DIC image below shows the terminal sporulation phenotype. Yellow and blue boxes indicate magnified regions of the kymograph. Scale bar: 2 µm. Additional examples of kymographs can be found in . ( E ) Fluorescence intensity traces of FtsZ-YPet (Z-rings) over time derived from sporulating WT (SS12) and Δ sepH mutant hyphae (MB750). Shown are the mean fluorescence intensity traces (mean ± SEM) collected from Z-rings of five sporulating hyphae for each strain. ( F ) Width of Z-rings in sporulating hyphae of WT (SS12) and sepH -deficient hyphae (MB750). The same data set as in ( E ) was used and the mean width for reach replicate (colored dots, n = 5) ±95% CI was plotted. ( G ) HADA labeling of peptidoglycan to visualize cross-walls in WT, Δ sepH (SV56), and Δ sepH/sepH + (MB747) hyphae. Spores of each strain were germinated and grown in the presence of 0.25 mM HADA for 5 hr in a microfluidic device before imaging. Scale bar: 20 μm. Figure 2—source data 1. Time-lapse fluorescence image series of selected and straightened hyphae (SS12) used to generate kymograph shown in . Figure 2—source data 2. Time-lapse fluorescence image series of selected and straightened hyphae (MB750) used to generate kymograph shown in . Figure 2—source data 3. Custom Python and R-scripts and extracted fluorescence intensities of Z-rings used to generate .

Article Snippet: For HADA (7-hydroxycoumarin 3-carboxylic acid-amino-D-alanine) labeling , spores were loaded into BA04 microfluidic plates (CellASIC ONIX).

Techniques: Mutagenesis, Fluorescence, Expressing, Derivative Assay, Labeling, Imaging

Journal: eLife

Article Title: A conserved cell division protein directly regulates FtsZ dynamics in filamentous and unicellular actinobacteria

doi: 10.7554/eLife.63387

Figure Lengend Snippet:

Article Snippet: For HADA (7-hydroxycoumarin 3-carboxylic acid-amino-D-alanine) labeling , spores were loaded into BA04 microfluidic plates (CellASIC ONIX).

Techniques: Amplification, Western Blot, Recombinant, Plasmid Preparation, Purification, Sequencing, Transformation Assay, Time-lapse Microscopy, Sedimentation, Staining, Synthesized, Software