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. 2012 Dec 7;42(2):460–468. doi: 10.3892/ijo.2012.1730

Figure 5.

Figure 5.

Effects of siRNA knockdown of RhoGDIβ in HeLa cells. (A) Stealth RNAi duplexes were transfected into HeLa cells and the protein levels of RhoGDIβ and RhoGDIα were determined by immunoblot analysis using anti-RhoGDIβ (556498) or anti-RhoGDIα/RhoGDIβ (66586E) antibodies at the indicated time after transfection. To estimate the degree of knockdown serially diluted samples of negative control were immunoblotted. Similar results were obtained from three independent experiments. (B and C) Cells were fixed with methanol 72 h after transfection of Stealth RNAi duplexes and stained with anti-γ-tubulin antibody (red) and DAPI (blue). The number of abnormal metaphases, such as (B) monopolar spindle and (C) multiple centrosomes, was counted under a fluorescence microscope. The total numbers of metaphases observed were 176, 229, 276 and 233 for the negative control, nos. 153, 469 and 800, respectively. Similar results were obtained from two independent experiments. (D) Representative images of aberrant position and number of centrosomes observed in RhoGDIβ knockdown cells. (E) DNA histograms were obtained by flow cytometry 72 h after transfection of Stealth RNAi duplexes. Similar results were obtained from three independent experiments.