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. 2011 Jun 22;6(6):e21123. doi: 10.1371/journal.pone.0021123

Figure 3. Colocalization of punctate vRNP signals with Rab11.

Figure 3

(A–C) Localizations of cytoplasmic vRNPs and transiently expressed human Rab proteins. Influenza A virus was infected to MDCK cells transiently expressing AcGFP-tagged human Rab11A, Rab25, and Rab17 (panels A–C, respectively). At 7 hpi, vRNPs were immunostained with mAb61A5 (center image in each set) and visualized by confocal microscopy with AcGFP-Rab proteins (right images). Enlarged images of indicated areas (white boxed) were also shown (lower images). Scale bars are 10 and 5 µm (upper and lower images, respectively). (D) Localizations of transiently expressed human Rab11A, Rab25, and Rab17. FLAG-Rab25 (upper) and FLAG-Rab17 (lower) (center images) were coexpressed with AcGFP-Rab11A (right images) in MDCK cells. Nuclei were stained with DAPI (blue, left images). Scale bar is 5 µm. (E and F) Colocalization of vRNP with endogenous Rab11. Progeny vRNPs were similarly stained with mAb61A5. Endogenous canine Rab11 (right images) was visualized with rabbit anti-Rab11 polyclonal antibody. (E) XY presentation. Scale bars are 40 and 5 µm (upper and lower images, respectively). (F) XZ presentation. Z-stacks of confocal images were acquired at 0.5 µm z-axis interval. Z-projection of maximum intensities (top image) and reconstitution of a xz plane (lower 3 images) were processed by using ImageJ software. Dotted line indicates the position of the reconstituted xz plane. Scale bar is 10 µm.