Figure 2: Trout gill bacteria are predominantly coated with IgT. | Nature Communications

Figure 2: Trout gill bacteria are predominantly coated with IgT.

From: Mucosal immunoglobulins at respiratory surfaces mark an ancient association that predates the emergence of tetrapods

Figure 2

(a) Representative flow cytometry histograms showing the staining of gill bacteria with IgT, IgM and IgD. Bacteria were stained with anti-trout IgT (green line), anti-trout IgM (red line) or anti-trout IgD (magenta line) mAbs or isotype controls (shaded histograms). (b,c) Percentage of gill bacteria coated with IgT, IgM or IgD (b) or coated with IgT and IgM, IgT and IgD, IgM and IgD, IgT and IgM and IgD (n=16) (c). The median percentage is shown by a red line. Statistical differences were evaluated by the nonparametric Mann–Whitney test. (d) Differential interference contrast (DIC) images of gill bacteria stained with a DAPI-Hoeschst solution (blue), anti-IgT (green), anti-IgM (red) or anti-IgD (magenta), and merging IgT, IgM and IgD stainings (Merge). (Isotype-matched control antibody staining is shown in Supplementary Fig. 3 online). (Scale bars, 5 μm). (e) Immunoblot analysis of IgT, IgM and IgD on gill bacteria. Lane 1, 0.1 μg of purified IgT, IgM or IgD; lanes 2–7, gill bacteria (n=6 fish). (f) Percentage of total gill mucus IgT, IgM or IgD coating gill bacteria (n=13). The median is shown by a red line. Statistical differences were evaluated by one-way ANOVA with Bonferroni correction. Data are representative of at least three independent experiments. ANOVA, analysis of variance.

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