Figure 9—figure supplement 3. | A long-term epigenetic memory switch controls bacterial virulence bimodality

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A long-term epigenetic memory switch controls bacterial virulence bimodality

Figure 9—figure supplement 3.

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The Hebrew University of Jerusalem, Israel
Figure 9—figure supplement 3.
Download figureOpen in new tabFigure 9—figure supplement 3. Invasion of HeLa cells by EPEC from SMALL and BIG colonies.

(A) Schematic illustration of gentamicin protection assay of HeLa cells infected with a 1:1 mixture of wild-type EPEC from SMALL and BIG colonies tagged with GFP and mCherry, respectively. (B) Plot of the results of the gentamicin protection assay for wild-type EPEC bacteria and for the ΔperA mutant. Data were normalized to initial CFU (t = 0 min) and are presented as the means ± s.d. of at least three technical repeats. (C) Schematic illustration of the microscopy-based gentamicin invasion assay together with induction dynamics. Induction of GFP fluorescence by IPTG 1 hr after gentamcin treatment enables the visualization of only live intracellular bacteria. (D) Confocal images (top and side views) of intracellular bacteria expressing GFP using the protocol described in (C). HeLa cells were fixed and stained for actin (red) and DNA (blue). Scale bar: 10 μm. (E) Time-lapse microscopy of HeLa cells infected according to the protocol shown in (C) showing growth of intracellular bacteria. Overlay of phase contrast and green fluorescence images are shown. Similar results were obtained in at least three different locations and in three independent biological replicates. Scale bar: 25 μm.