Video 4.
Klp10A regulates the length of the centriolar MTs. Movies show electron tomograms of centrioles in longitudinal orientation in WT (left) and Klp10A (right) third instar larval wing disc cells. Tilt series of 150-nm thick sections were collected from −55° to 55° with 1° angular increment in a transmission electron microscope (TECNAI T12; FEI) at 13,000× using SerialEM (Mastronarde, 2005). Tomograms were reconstructed by R-weighted back projection with the user interphase eTomo and rendered three-dimensional using the IMOD package (Kremer et al., 1996). Centrioles were oriented along their longitudinal axis. Each movie frame in the tomogram is a projection of 10 individual slices. Note that the WT electron tomogram is the same as that shown in Video 1, and it is shown again here simply for ease of comparison. Centriolar MTs are dramatically elongated in the Klp10A mutant centriole, and these extensions consist largely of doublet MTs. Bar, 100 nm.