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. Author manuscript; available in PMC: 2015 Feb 9.
Published in final edited form as: Nat Commun. 2014 Oct 22;5:5205. doi: 10.1038/ncomms6205

Figure 2. Trout nasal bacteria are coated by secretory immunoglobulins.

Figure 2

(a) Differential interference contrast images of trout olfactory organ stained with NONEUB oligoprobe by fluorescence in situ hybridization. Note that no bacterial staining was observed (N = 5). (b) Differential interference contrast images of trout olfactory organ stained with EUB338 oligoprobe (magenta) that stains ~90% of all eubacteria. Abundant bacteria (magenta) can be observed in the lumen of the olfactory organ and some within the olfactory epithelium (N = 5). Nuclei (blue) are stained with the DNA-intercalating dye DAPI. Scale bar, 20 μm. (c–e) Fluorescent microscopy images of trout nasal bacteria stained with a DAPI-Hoeschst solution (blue; c), anti-IgM (red; d) or anti-IgT (green; e). Orange arrows indicate bacteria that are positive for DAPI, IgM and IgT (double-coated population). Scale bar, 20 μm. (f) Immunoblot analysis of IgTand IgM on nasal-associated bacteria and gut luminal bacteria (as a positive control). (g) Percentage of trout nasal-associated bacteria that are uncoated, coated with IgT, IgM or both IgTand IgM quantified from immunofluorescence microscopy images. (h) Percentage of trout nasal-associated bacteria that are uncoated or coated with at least one antibody isotype quantified from immunofluorescence microscopy images. Data (mean±s.e.) are representative of three independent experiments (N = 6).