Fig. 5: H2A entry into the bacterial cytoplasm inhibits growth, perturbs chromosomal organization, and suppresses transcription.

a Growth profiles of E. coli that were untreated, electroporated in the absence of any treatment (empty electroporation), treated with 10 μg/mL AF-H2A, 2 μM LL-37, or both, or electroporated with 10 μg/mL AF-H2A and cultured with the same concentration of AF-H2A. Cells were cultured in minimal medium containing 1 mM magnesium. AF-H2A was mixed with unlabeled H2A (1% AF-H2A, combined concentration of 10 μg/mL) (n = 4 for each condition). b CFUs per microliter of cultures that were treated for 1 h under identical conditions as (a) and plated on non-selective LB plates (n = 4 for each condition). c AF-H2A fluorescence intensities of E. coli that were cultured under identical conditions as (a) (n = 3 for each condition). d Representative phase contrast, SYTOX fluorescence, and merged images of E. coli that were untreated or treated with 10 μg/mL H2A, 2 μM LL-37, or both. The fluorescence images are displayed using the full range of pixel values in the images. e Corresponding principal component analysis (PCA) of images of SYTOX-stained E. coli (n = 3 for each condition). Each cell is represented as a point on a PCA plot, which is then transformed into a density plot. The color scales indicate normalized cell densities. f Representative phase contrast, HupA-mRuby2 fluorescence, and merged images of E. coli that express HupA-mRuby2 that were untreated or treated with 10 μg/mL H2A, 2 μM LL-37, or both. g Corresponding PCA analysis of E. coli expressing HupA-mRuby2 that has been transformed to density plots (n = 3 for each condition). For imaging, cells were immobilized for 3 h on agarose pads containing 2 μM LL-37, 10 μg/mL H2A, or both. For SYTOX analysis, pads additionally contained 5 μM SYTOX. Bars and points are shown as mean ± SEM of biologically-independent experiments. One-way ANOVAs were performed in (b) and a two-way ANOVA was performed in (c) in which the 30-min data were excluded the analysis. See Supplementary information for raw statistical data. Images are representative of three independent experiments. Scale bars represent 2 μm.