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Figure 8


Fig. 8. Depletion of myosin VI and LMTK2 leads to reduction in EHD1- and EHD3-positive tubule formation. (A,B) HeLa cells stably expressing (A) GFP-EHD1 or (B) GFP-EHD3 were treated with siRNA targeting myosin VI or LMTK2, fixed and mounted for immunofluorescence. Representative examples of (A) GFP-EHD1 or (B) GFP-EHD3 distribution are shown for each KD and control cell. (C) To quantify the number of EHD3 positive tubules in KD and control cells at least 200 randomly chosen cells were counted for each condition and scored as having either no, few (<15) or multiple (>15) tubules. The results are given as percentage of cells for each category and are expressed as the mean ± s.d. from three independent experiments, each performed in triplicate. (D,E) The number of tubules was counted and the length of individual tubules was measured in 25 cells for each knockdown. Cells without tubules were not taken into account. Branches were considered as separate tubules. The histograms depict the (D) number of tubules per cell and the (E) length of individual tubules. In (A) confocal z-stacks and in (B) wide-field images are shown. Bars, 10 µm.





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