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To potentially increase access for the antibodies, we used isolated cell envelopes for our labeling experiments, but again failed to observe labeling of the PilQ nozzles at cell-cell junctions

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Cells were stained for sgRNA expression with an APC-conjugated anti-mouse Thy1.1 antibody (202526, BioLegend, 1:400) and human TruStain FcX receptor blocking solution (422302, BioLegend, 1:1000) for 5 min at 4C in FACS buffer (PBS with 5% FBS and 1mM EDTA)