Fig. 4: Luminal EVs derived from AmII-associated germ-free mice inhibit AmI growth in vitro. | Nature Communications

Fig. 4: Luminal EVs derived from AmII-associated germ-free mice inhibit AmI growth in vitro.

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

Fig. 4: Luminal EVs derived from AmII-associated germ-free mice inhibit AmI growth in vitro.The alternative text for this image may have been generated using AI.

A, B Levels of EB-AMDK19 (AmI) (A) and EB-AMDK39 (AmII) (B) in feces from mono-associated GF mice at different time points. GF mice were gavaged with each clade and fecal DNA was prepared to determine luminal levels of each clade by qPCR (n = 4). C, D Changes in growth of EB-AMDK19 (C) and EB-AMDK39 (D) in vitro upon treatment of whole fecal soluble fractions (SF) from GF mice, and whole fecal SF, heat-inactivated whole fecal SF, and size-fractionated SF with indicated cut-off from EB-AMDK39-associated GF mice. Growth was assessed after 24 h upon treatment. 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 Identification of inhibitory size-exclusion fractions derived from fecal SF of GF (left) and EB-AMDK39-associated GF mice (right). Fractions prepared by size-exclusion chromatography were used to treat EB-AMDK19 and EB-AMDK39 culture at a final concentration of 20% (v/v). Growth of each clade at 37 °C for 24 h is depicted as a percentage of media control. Points and connecting lines were drawn based on the clade growth patterns (% of medium control) according to the fractions. Growth of EB-AMDK19 and EB-AMDK39 was marked in black and blue, respectively. The column bar graph was drawn using the protein concentration (mg/mL) in each fraction (red). Data represent the averages of triplicate measurements. Two independent experiments showed similar results. Statistical differences were determined by two-way ANOVA with Sidak’s multiple comparison tests. Error bars represent SEM. F Disk diffusion test to show changes in growth of EB-AMDK19 upon treatment with enriched EVs from EB-AMDK39 in vitro culture, fecal SF derived from GF (B), and EB-AMDK39-asssociated GF mice (C). Source data are provided as a Source Data file.

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