Figure 5.

Immuno-EM localization of Cep164. (A) U2OS cells were subjected to preembedding immuno-EM. Cells were labeled with anti-Cep164 antibody R171 followed by Nanogold-coupled secondary antibody. Note that Cep164 localizes to very distal appendage structures on parental centrioles (arrowheads, also see B), and sometimes electron-dense material (presumably reflecting subdistal appendages) can be seen proximal to the Cep164 staining (arrows in second panel and in B [top]). The schematic diagrams (dotted lines) in the two top panels illustrate the positions of “mother” (MC) and “daughter” centrioles (DC), along with the presumed positions of distal and subdistal appendages. (B and C) U2OS cells were transfected for 48 h with control (GL2) or Cep164-specific siRNA duplexes and subjected to preembedding immuno-EM. Cells were labeled with either anti-Cep164 or anti-ninein antibody followed by Nanogold-coupled secondary antibody. Bars, 250 nm.