Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2013 Feb 1;24(3):222–233. doi: 10.1091/mbc.E12-03-0198

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2013 Kienzle et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

“ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.

PMC Copyright notice

FIGURE 5: — Depletion of PKD affects the cleavage of the noncompact zones in G2. (A) HeLa cells cultured on collagen-coated coverslips were synchronized at the G1/S border and transfected with siLacZ or siPKD1/2. One sample of each setup was additionally transfected with a caMEK1- plasmid fused to mCherry. Cells were released for 10, 12, and 14 h, fixed, and stained with antibodies specific for GM130 and pH3. Left, cells in late G2 were scored for their Golgi complex morphology. The graph displays the results of the quantification of cells in G2 stage harboring an intact Golgi complex (percentage of total G2 cells). At least 35 cells per sample and experiment were analyzed. Shown is mean ± SEM of three independent experiments. Statistical evaluation was performed using one-way ANOVA followed by Tukey’s multiple comparison test. **p < 0.01. Right, representative images showing Golgi complex morphology of control cells (siLacZ), PKD1/2-depleted cells (siPKD1/2), and siRNA-transfected cells expressing caMEK1-mCherry (siLacZ + caMEK1 and siPKD1/2 + caMEK1) in G2 stage. Color code: GM130 (green), pH3 (cyan), caMEK1-mCherry (red). Scale bar, 10 μm. (B, C) HeLa cells stably transfected with ManII-GFP and transiently transfected with siLacZ or siPKD1/2 were left untreated (interphase) or treated with bisbenzimide (G2 block). The region marked with a rectangle was photobleached, and FRAP was measured over time. (B) Representative images illustrating the Golgi complex before (prebleach), directly after (bleach). and during recovery (postbleach). The bleached areas are delineated by white-bordered rectangles. Scale bar, 10 μm. (C) The curves show the normalized FRAP kinetics of interphase cells and G2 cells over time. Fluorescence recovery in bleached areas was monitored every 2 s. Curves of fluorescence intensities were normalized to nonbleached areas. Shown is mean ± SEM of two independent experiments (10 cells/sample per experiment). (D) Y_max (maximum fluorescence recovery) was determined by fitting the curves shown in C to a one-phase exponential equation.