Fig 5.
LcrVS324 caps YscF needles that lack YopD. (A) Cultures of Y. enterocolitica CT1 (ΔlcrV) harboring plcrVW22703, plcrVS228, plcrVS324, or no plasmid were centrifuged, and the bacterial sediment was sheared to break off type III needle complexes. Bacteria were removed by slow-speed centrifugation, and the filtered supernatant was ultracentrifuged to sediment type III needle complexes, which were analyzed by transmission electron microscopy. The arrowheads identify LcrV caps of type III needle complexes. (B) Y. enterocolitica CT1 (ΔlcrV) and Y. enterocolitica ΔpYVe227 cultures were subjected to the isolation of type III needle complexes as described above. Samples were analyzed by immunoblotting with rabbit antisera raised against LcrV (αLcrV), YopD (αYopD), and YscF (αYscF).