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Figure 6


Fig. 6. Analysis of EML3 function in spindle formation. Knockdown of EML3 was performed as described in Fig. 5 using two different siRNA oligonucleotides specific to EML3 (1 and 2) and cells fixed with paraformaldehyde 80 hours post transfection. (A) Mitotic indices after gene knockdown were determined from 480 cells and means and s.d. from three independent experiments were plotted (left panel). The sums of cells at the different mitotic stages after gene knockdown were determined in the mitotic cells (right panel). (B) Representative immunofluorescence images of aberrant metaphase-like structures with unaligned chromosomes. Merge: DNA (blue) and Tubulin (green). Scale bars: 10 µm. Quantification of phenotypes resulting from EML3 knockdown is shown on the right. Means ± s.d. from three independent experiments (n=100) were plotted. (C) The maximum extension of metaphase chromosomes in a virtual pole-to-pole axis of the mitotic spindle was determined in 20 images after treatment with the respective siRNA oligos as indicated. Single values (left) and average values and s.d. (right) were plotted.





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