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. 2014 Feb 11;5(1):e01024-13. doi: 10.1128/mBio.01024-13

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Copyright © 2014 Miller et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

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FIG 6 — Mutations in PilE_SB(Z2491 D-region) and PilE_SB(Z5463 D-region) restore ezrin recruitment. (A and B) Sequence alignment of D-regions from pilin variants SB, Z2491 and Z2491_{ANGKQ→KTDDK}, Z5463, and Z5463_ND→TTA. Mutated residues are circled in red. (C to F) HDMECs (C and E) and FaDu cells (D and F) were infected with strains of N. meningitidis expressing PilE_SB(Z2491 D-region) and PilE_SB(Z2491 D-region)_{ANGKQ→KTDDK}, PilE_SB(Z5463 D-region) and PilE_SB(Z5463 D-region)_ND→TTA. HDMECs were infected with the PilV⁻ PilT⁻ derivatives of these strains. (C and D) The ezrin recruitment index was estimated by determining the proportion of colonies that efficiently recruit ezrin at the site of adhesion and expressed as normalized mean values (±SEM) of three independent experiments in duplicate. *, P < 0.002 (Student’s t test). (E and F) Ezrin was immunostained (in green), and DNA was stained using DAPI (in blue). Microcolonies are shown by arrows. Scale bars, 10 µm.