Fig. 3: Extracellular vesicles derived from AmII inhibit AmI growth in vitro. | Nature Communications

Fig. 3: Extracellular vesicles derived from AmII inhibit AmI growth in vitro.

From: Clade-specific extracellular vesicles from Akkermansia muciniphila mediate competitive colonization via direct inhibition and immune stimulation

Fig. 3: Extracellular vesicles derived from AmII inhibit AmI growth in vitro.The alternative text for this image may have been generated using AI.

A Changes in EB-AMDK19 (AmI) growth upon treatment with unprocessed, heat-inactivated (HI) and protease-treated EB-AMDK39 (AmII)-derived culture supernatant. AmII-derived supernatants were heat-inactivated at 95 °C for 15 min or treated with proteinase K (1 mg/mL) for 16 h at 37 °C. Proteinase K-treated samples were supplemented with protease inhibitor (1 mM). B EB-AMDK19 growth inhibition by EB-AMDK39-derived culture supernatant fractionated by molecular weight. EB-AMDK19 growth was assessed after 24 h in the presence of 20% size-fractionated EB-AMDK39-derived culture supernatant. C Identification of inhibitory size-exclusion fractions derived from EB-AMDK39-derived culture supernatant. Fractions prepared by size-exclusion chromatography were treated into EB-AMDK19 culture at a final concentration of 10% (v/v). Growth of EB-AMDK19 at 37 °C for 24 h was depicted as a percentage of media control. Points and connecting lines are drawn to show the EB-AMDK19 growth changes according to the fractions. Column bar graph shows the protein concentration (mg/mL) in each fraction. D Dose dependent inhibition of EB-AMDK19 growth by enriched EB-AMDK39-derived EVs. EV particle number was determined by nano particle tracking analysis. Enriched EB-AMDK39-derived EVs at various EV-to-cell ratios were used to treat EB-AMDK19 culture (1 × 106 CFU/culture) and growth was assessed after 24 h. A–D Growth is depicted as a percentage of media control (n = 3 biological replicates). At least, two independent experiments showed similar results. Statistical differences were determined by one-way ANOVA with Tukey’s multiple comparison tests. ns: not significant. Error bars represent SEM. E Disk diffusion test to monitor the growth inhibition of each clades by enriched EVs derived from EB-AMDK19 (EVs-EB-AMDK19) and EB-AMDK39 (EVs-EB-AMDK39). F Transmission electron micrographs showing enriched EVs derived from EB-AMDK19 or EB-AMDK19 and the structural integrity of each clade strain upon treatment with enriched EVs derived from EB-AMDK19 or EB-AMDK19. Bars indicate 500 nm. At least two independent experiments showed similar results. Source data are provided as a Source Data file.

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