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. 2016 Feb 9;11(2):e0148887. doi: 10.1371/journal.pone.0148887

Fig 7. Effect of bile exposure on B. fragilis OMV formation.

Fig 7

(A) Growth of B. fragilis YCH46 in GAM supplemented with the indicated bile concentrations. (B) qPCR was used to determine the relative abundance of the IVp-I-ON or IVp-II-ON genotypes after exposure to the indicated bile concentration. (C) Effect of bile exposure on the expression of an IVp-I-regulated EPS production gene (BF2767) and IVp-II-regulated membrane protein genes (BF3397 and BF3403). (D) OMV production of B. fragilis increased 7-fold after exposure to 5% bile. (E) Schematic of the 3 x FLAG-tag fusion to the C-terminus of the BF3397-encoded protein. (F) Enhancement of the BF3397-encoded membrane protein in response to 5% bile. A B. fragilis YCH46 derivative with a 3 x FLAG-tag sequence fused to BF3397, as shown in panel E, was exposed to 5% bile: the production of BF3397 was assessed by Western blotting with an anti-FLAG antibody. Upper, middle, and lower bands indicate the anti-FLAG tag antibody used for immunoprecipitation, BF3397 (unprocessed form), and N-terminally processed BF3397 (indicated by arrowheads), respectively.