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. 2022 Jun 23;13:3603. doi: 10.1038/s41467-022-31176-9

Fig. 2. Intracellular and extracellular forms of Ot are metabolically distinct.

Fig. 2

A Confocal microscopy image of an L929 cell infected with Ot for 5 days and labeled with the clickable methionine analog homopropargylglycine (HPG, green) and an antibody against surface protein TSA56 (red). DNA (Hoechst) is shown in blue. The host cell outline is showed by a dashed line, and an arrow indicates a metabolically inactive bacterium located outside the host cell. B Quantification of metabolic activity, as measured by a detectable HPG signal, in IB and EB Ot. L929 cells were infected with Ot for 5 days, fixed, and labeled with TSA56 and HPG. Bacteria were identified as being IB or EB, based their position relative to host cells, and the percentage of each population that was HPG-positive was quantified. The percentage positivity for ten randomly imaged field is shown here, and this data is representative of >3 independent repeats of this experiment. Groups were compared using a two-sided Mann–Whitney test. Statistical significance for all data in this figure was determined using Graphpad Prism software. *p value ≤ 0.05, **p value ≤ 0.01, ***p value ≤ 0.001, ****p value ≤ 0.0001, ns not significant.