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. 2015 Dec 4;468(1-2):39–45. doi: 10.1016/j.bbrc.2015.10.169

Fig. 4.

Fig. 4

Wdr8 depletion results in shorter and tilted mitotic spindles. (A) Representative images of mitotic spindle morphologies of HeLa cells treated with control or Wdr8 siRNA are shown. Immunofluorescence microscopy was performed with antibodies against α-tubulin (red) and γ-tubulin (green). (B) Quantification of spindle length. Data derived from (A) is shown. At least 150 cells were counted (n = 3). ****p < 0.0001. (C, D) Schematic presentation of spindle angle (C, θ represents angle °) and quantification of spindle orientation (D) are shown. Data derived from (A) is shown. At least 150 cells were counted (n = 3). **p < 0.01. (E) Rescue of defective spindle morphologies by introduction of siRNA-resistant Wdr8 (Wdr8*) constructs. HeLa cells depleted of Wdr8 were transfected with plasmids producing siRNA-resistant Wdr8 and immunofluorescence microscopy was performed as in (A). Images of Venus (Venus only on the top and Venus-Wdr8 on the bottom) are shown. Note that Venus-Wdr8 localises to the centrosomal area (bottom). Quantification data is presented in the right-hand side. At least 150 cells were counted (n = 3). ****p < 0.0001. Scale bars, 5 μm.