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To test the hypothesis that microbiota influences Ace2 expression, we compared two groups of concomitantly raised gnotobiotic (germ-free, GF) rats with one group acquiring gut microbiota through co-housing for 10 days with conventional specific pathogen-free rats. 16S rRNA sequencing analysis of fecal samples confirmed successful colonization of 9 bacterial phyla in the GFC rats (Figure [A]). Reconstitution of the gut microbiota for 10 days markedly decreased the colonic Ace2 expression in GFC rats, compared with GF rats (Figure B). As the only variable between the 2 groups was the presence or absence of the microbiota, our observation indicates that the gut microbiota colonized in the GFC rats caused the decrease in colonic Ace2 expression.