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. 2016 Oct 4;2:16033. doi: 10.1038/celldisc.2016.33

Figure 2.

Figure 2

Depletion of ASK1 impairs spindle orientation and positioning. (a) Immunoblots showing the expression of ASK1 and α-tubulin in control or ASK1 siRNA-treated HeLa cells. (b) Scheme for spindle angle (α) measurement. (c–f) Immunofluorescence images (c), spindle length (d), spindle angle (e) and spindle angle distribution (f) of metaphase HeLa cells transfected with control or ASK1 siRNAs and stained with anti-α-tubulin (green) and anti-γ-tubulin (red) antibodies and DAPI (blue). The position of the z stage is indicated in micrometers; 3D, xy projection. n=30 cells per group. Scale bars, 5 μm. (g) Scheme for spindle displacement distance (d) measurement. (h–k) Immunofluorescence/phase-contrast images (h), cell diameter (i), spindle displacement distance (j) and spindle displacement distance distribution (k) of control and ASK1 siRNA-treated metaphase HeLa cells stained with anti-α-tubulin antibody (green) and DAPI (blue). n=30 cells per group. Scale bars, 5 μm. Experiments were performed three times. Values are mean±s.e.m. *P<0.05, **P<0.01, ***P<0.001; NS, not significant.