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. 2019 Oct 23;203(11):3054–3067. doi: 10.4049/jimmunol.1900863

FIGURE 2.

FIGURE 2.

A majority of trout pharyngeal bacteria are predominantly coated with IgT. (A) Representative flow cytometry histograms showing the staining of pharyngeal bacteria with IgT, IgM, and IgD. Bacteria were stained with isotype controls (shaded histograms), anti-trout IgT (green line), anti-trout IgM (red line), or anti-trout IgD (magenta line) mAbs, respectively. (B) Percentages of pharyngeal bacteria coated with IgT, IgM, or IgD (n = 25). The median percentage is showed by a red line. (C) Immunoblot analysis of IgT, IgM, or IgD coating on pharyngeal bacteria. Lane 1, 0.1 μg of purified IgT, IgM, or IgD; lanes 2–7, pharyngeal bacteria (n = 6). (D) Percentages of total pharyngeal mucus IgT, IgM, or IgD coating pharyngeal bacteria (n = 12). The median is shown by a red line. The statistical differences in (B) and (D) were evaluated by one-way ANOVA with Bonferroni correction. Data are representative of at least three independent experiments. (E) Differential interference contrast (DIC) images of pharyngeal bacteria stained with a DAPI/Hoechst solution (blue), anti-IgT (green), anti-IgM (red), or anti-IgD (magenta) and merging IgT, IgM, and IgD staining (Merge) (isotype-matched control Ab staining, Supplemental Fig. 2A). Scale bar, 5 μm.