Supplementary Figure 2: C2CD3 interacts with the BBSome and localizes to centriolar satellites.
From: The oral-facial-digital syndrome gene C2CD3 encodes a positive regulator of centriole elongation
(a) C2CD3 coimmunoprecipitates with most BBSome subunits. Plasmids encoding GFP-C2CD3 and a Myc-tagged BBSome subunit were cotransfected in HEK cells, and complexes were precipitated with anti-Myc antibody. Inputs and eluates were resolved by SDS-PAGE and immunoblotted for Myc or GFP. For the Myc immunoblot, 2.5 equivalents of eluate and 1 equivalent of lysate were run on the gel. Both Myc immunoblot panels are cropped from the same film exposure. For the GFP immunoblot, 25 equivalents of eluate and 1 equivalent of lysate were run on the gel. Both GFP immunoblot panels are cropped from the same film exposure. (b) C2CD3 localizes to centriolar satellites. IMCD3-[GFP-C2CD3] cells were stained for PCM-1 (red) and DNA (blue). Most foci of C2CD3 signal overlap with PCM-1 except for two spots in the center of the cell (pointed by arrowheads within the magnified region in the inset). Scale bars, 5 μm; the inset is 2 μm by 2 μm. (c) The distribution and overall number of centriolar satellites are not affected by the loss of C2CD3. Heterozygous MEFs or mutant MEFs were stained for PCM-1 (green) and DNA (blue). Scale bar, 5 μm.
