Fig. 5

Mitotic phenotypes caused by co-depletion of Patronin and either Asp, Klp10A or Dgt6. (a) Examples of the mitotic phenotypes of cells depleted of Patronin, Klp10A, Asp, or Dgt6 and co-depleted of Patronin and Klp10A, Patronin and Asp, or Patronin and Dgt6. Cells were stained for DNA (DAPI, blue), α-tubulin (green) and the centrosomal marker Spd2 (red). Note the short collapsed spindles in Patronin-depleted cells and the long “rescued” spindles in cells co-depleted for either Patronin and Klp10A or Patronin and Asp. See text and Table 1 for detailed descriptions of the observed phenotypes. Scale bar, 5 μm. (b) RT-qPCR results showing that RNAi against Patronin, asp, Klp10A, or dgt6 strongly reduces the level of the corresponding transcripts relative to a mock control that is set to 100%. For each gene, the reduction has been calculated by averaging the transcript levels detected in at least three independent RNAi experiments; RpL32 was used as an endogenous reference gene. (c) Box and Whisker plots showing the quartile ranges of metaphase spindle length in control cells and cells depleted of Patronin, Asp, Klp10A, both Patronin and Asp, and both Patronin and Klp10A. Patronin-depleted cells have spindles significantly shorter than control spindles (p < 0.01), while Asp-depleted and Patronin and Asp co-depleted cells have spindles significantly longer than those of control cells (p < 0.01). Also note that Klp10A-depleted and Patronin and Klp10A co-depleted cells have spindles significantly longer than those of control cells (p < 0.01)