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Fig. S1
Comparison of E. coli K12 strain with enteric adherent invasive E. coli strains in notochord infection model.
(A) Diagram showing the injection of Crimson expressing E. coli in the notochord (arrow: injection site) in triple transgenic larvae tg(RCN3:gal4/UAS:DsRed/mpx:GFP) at 2 days post-fertilization larva (dpf). (B) Larvae were analyzed by confocal microscopy at 5 hours post-injection (hpi) of either PBS or Crimson-E. coli. Notochord images are representative 3D projections of overlaid fluorescence channels: DsRed (blue), GFP (green) and Crimson (magenta). The right panel is a projection of cross-section view of the notochord in the region indicated by the dotted line. Dashed circle outlines the notochord. Scale bar: 30 μm. (C) Electron microscopy of the notochord region in infected larvae at 4 hpi. b: bacteria, nc: notochord, col: collagen sheath. Scale bar = 1μm. (D) GFP expressing Escherichia coli strains (K12, AIEC LF82, LF82-ΔlpfA and JM83-ΔmsbB) were injected in the notochord of tg(mpx:GFP) embryos at 2 dpf. GFP (E. coli and neutrophils) was analysed by fluorescence microscopy at 1 dpi. In AIEC LF82, LF82-ΔlpfA and JM83-ΔmsbB infections, bacteria were cleared and neutrophil recruitment to the notochord (N) was induced similarly to K12 infections (arrowheads). Scale bar: 400 μm. (E) Survival curves of zebrafish larvae that have been infected in their notochord with indicated Escherichia coli strains from 0 to 2 dpi (Log rank test, ns = not significant p>0.05, N is indicated on in the figure).