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. 2016 Apr 19;7:11182. doi: 10.1038/ncomms11182

Figure 5. In vivo analysis reveals that Tum/RacGAP is required for Pav/kinesin-6 localization.

Figure 5

(a) Left: western blot of Tum/RacGAP and Pav/kinesin-6 proteins in control and in Tum/RacGAP-depleted S2 cells. H3 protein used as a control. Right: Pav/kinesin-6 levels in Tum/RacGAP-depleted and control S2 cells. Images captured with the same exposure time. Scale bar, 5 μm (b) Left: auto-fluorescence test of secondary antibody used against Pav/kinesin-6. S2 cells stained with DAPI (blue), Tubulin (green) and secondary antibody (red) without anti-Pav/kinesin-6 primary antibody. Right: average fluorescence intensity of Pav/kinesin-6 normalized to the control in Tum/RacGAP-depleted cells during metaphase. (c) Metaphase, anaphase and telophase in control (left panel) and Tum/RacGAP-depleted (right panel) S2 cells. White arrowheads: localization of Pav/kinesin-6 at centrosomes of early-late anaphases. Red arrowheads: Pav/kinesin-6 localization on telophase centrosomes. Cells stained for tubulin (green), DNA (blue) and Pav/kinesin-6 (red). Scale bar, 5 μm. (d) Feo/PRC localization during metaphase, anaphase and telophase in control (left panel) and Tum/RacGAP-depleted (right panel) S2 cells. Cells stained for tubulin (green), DNA (blue) and Feo/PRC (red). Scale bar, 5 μm.