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. 2012 May 15;23(10):1838–1845. doi: 10.1091/mbc.E11-12-1043

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© 2012 Holland 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).

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FIGURE 4: — Repression of Plk4 by siRNA results in the loss of centrioles due to impaired duplication. (A) Representative immunofluorescence images showing the localization of endogenous Plk4 in late-telophase cells. Red, α-tubulin; green, Plk4; blue. Scale bar, 5 μm. (B) As in A, except that cells stably overexpressing Plk4-EYFP were used. Red, α-tubulin; green Plk4-EYFP; blue, DNA. Scale bar, 5 μm. (C) Representative immunofluorescence images showing the level of Plk4 at the centrosome 48 h after transfection of Plk4^+/− RPE1 cells with Plk4 or GAPDH siRNA. Red, γ-tubulin; Green, Plk4; Blue, DNA. Scale bar = 5 μm. Graph shows the quantitative immunofluorescence analysis of Plk4 protein levels at the centrosome 48 h after cells were transfected with Plk4 or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) siRNA. Note that only cells with at least one centrosome were quantified in this analysis, and thus the level of Plk4 repression is likely to be underestimated. Bars show the mean of >42 cells per condition from two independent experiments. Error bars represent the SEM. (D) The fraction of Plk4^+/− RPE1 cells with the indicated number of γ-tubulin foci 48 h after transfection with Plk4 or GAPDH siRNA. Bars show the mean of >180 cells per condition from three independent experiments. Error bars represent the SEM. (E) Representative immunofluorescence images show phenotypes observed in mitotic Plk4^+/− RPE1 cells 48 h after transfection with Plk4 or GAPDH siRNA. Red, α-tubulin; green, centrin; blue, DNA. Scale bar, 5 μm.