To analyse percentages of piliated cells, each HI strain was back-diluted and grown to OD600 0.1–0.5 in PY broth at 29°C 200 rpm. Cells were then stained with 2.0% phosphotungstic acid (PTA) on carbon formvar copper grids (Agar Scientific) and analysed for the presence/absence of a pilus structure, as described previously [26] (link).
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