Data underlying Chapter 4 of PhD thesis: Working toward establishing the link between foci, filaments and membrane abundance

This collection of microscopy datasets investigates how the essential phospholipid synthesis enzyme PlsB regulates membrane growth and organization in <em>Escherichia coli</em>. Fluorescence microscopy data capture PlsB-msGFP2 localization during spheroplast formation, osmotic shocks, FRAP experiments, and altered protein expression levels. These experiments reveal how PlsB dynamics change when fatty-acid synthesis is inhibited, the cell wall is weakened, or membrane area is perturbed. Additional widefield fluorescence imaging of PlsB point-mutant strains assesses how individual amino acid substitutions affect protein clustering and membrane association. Electron microscopy datasets complement these observations by examining ultrastructural context in minicells, thin-sectioned cells, and purified PlsB samples under different preparation conditions. Negative stain and cryo-TEM imaging allow evaluation of potential PlsB oligomerization and membrane interaction at high resolution. Together, the fluorescence and EM datasets provide multiscale insight into the spatial organization of PlsB and its coupling to membrane biogenesis. Furthermore, images are provided showing successful formation of vesicles which can in the future be used as vehicles for in vitro study of PlsB. A MATLAB analysis script supports quantitative extraction of cell geometry and PlsB foci statistics, enabling reproducible interpretation of these imaging data.