Download TABLE?S3, DOCX file, 0.1 MB. Copyright ? 2018 LaSarre et al. (A) Spectral analysis of WT (red) and mutant (blue) strains grown in anaerobic PMsuccYE in 20?mol s?1 m?2 light, confirming loss of carotenoid synthesis in the mutant. (Inset) Photograph of concentrated cells of each strain. (B) Microscopy images of WT and cells from cultures used in panel A. Download FIG?S2, TIF file, 0.8 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S3? ICM localization pattern is conserved in non-UPP-bearing WT cells. (A) Lengths of cells containing one or two ICMs. For 1 ICM, 6,493?cells; for 2 ICMs, 648?cells. (B) Demograph of BChl fluorescence intensities measured along the medial cell axis of all cells in panel A. Cells were sorted from shortest to longest. No cell polarity was assigned. (C) Longitudinal distance (ICM spacing) GIBH-130 between ICMs within cells containing two ICMs plotted as a function of cell length. 648 ICM pairs. Download FIG?S3, TIF file, 0.9 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S4? Restricted ICM localization is conserved among diverse ecotypes of isolates grown in Rabbit polyclonal to Neuron-specific class III beta Tubulin anaerobic PMsuccYE in 8?mol s?1 m?2 light. Image contrast and brightness are not equivalent between fluorescence panels. Download FIG?S4, TIF file, 0.8 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms GIBH-130 of the Creative Commons Attribution 4.0 International license. FIG?S5? ICM localization is conserved across a range of light intensities. Shown are microscopy images of CGA009 grown in anaerobic PMsuccYE at different light intensities. Images are from among those used for analysis in Fig.?1E. Image contrast and brightness are not equivalent between fluorescence panels. Download FIG?S5, TIF file, 0.7 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S6? Fluorescence microscopy images of PNSB, with phase-contrast and overlay panels included. All species were grown phototrophically in 8?mol s?1 m?2 light. Image contrast and brightness are not equivalent between fluorescence panels. Download FIG?S6, TIF file, 2.2 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S1? Characteristics of select purple nonsulfur bacteria. Download TABLE?S1, DOCX file, 0.1 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S7? Quantitative analysis of ICM localization in adhesin-bearing and cells. (A) Microscopy image of cells showing BChl fluorescence (cyan) and polar adhesin stained with ConA-488 (false-colored red). Scale bars, 2?m. Image contrast and brightness are not equivalent between fluorescence panels. (B) Lengths of cells containing one or two ICMs. For 496?cells, and for 2 ICMs, 218?cells; for 846?cells, and for 2 ICMs, 155?cells. (C) Longitudinal distance (ICM spacing) between ICMs within cells containing two ICMs plotted as a function of cell length. For 218 ICM pairs; for 155 ICM pairs. Download FIG?S7, TIF file, 0.5 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S2? Bacterial strains, plasmids, and primers used in this study. Download TABLE?S2, DOCX file, 0.1 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. TABLE?S3? Alternative microscopes and filter sets for visualizing BChl-fluorescence. Download TABLE?S3, DOCX file, 0.1 MB. Copyright ? 2018 LaSarre et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 GIBH-130 International license. ABSTRACT In bacteria and eukaryotes alike, proper cellular physiology relies on robust subcellular organization. For the phototrophic purple nonsulfur bacteria (PNSB), this organization entails the use of a light-harvesting, membrane-bound compartment known as the intracytoplasmic membrane (ICM). Here we show that ICMs are spatially and temporally localized in diverse patterns among PNSB. We visualized ICMs in live cells of 14 PNSB species across nine genera by exploiting the natural autofluorescence of the photosynthetic pigment bacteriochlorophyll (BChl). We then quantitatively characterized ICM localization using automated computational analysis of BChl fluorescence patterns within single cells across the population. We revealed that while many PNSB elaborate ICMs along the entirety of the cell, species across as least two genera restrict ICMs to discrete, nonrandom sites near cell poles in a manner coordinated with cell.