Talpid3 interacts with PCM1 and regulates localization of centriolar satellites markers. (A) Western blotting of endogenous PCM1, Cep290, Talpid3, Cep97, and CP110 after immunoprecipitation of RPE1 cell extracts with antibodies indicated at the top of panel. IN, input. (B, D, and F) RPE1 cells transiently transfected with control (siNS), Talpid3, or Cep290 siRNAs were induced to quiescence for 48 h and processed for immunofluorescence with anti-glutamylated tubulin (GT335, green) and (B) anti-Cep290, (D) anti-PCM1, or (F) anti-BBS4 (red) antibodies. Bars, 5 µm. DNA was stained with DAPI (blue). (C and E) The percentage of quiescent RPE1 cells with accumulations of Cep290 (C) or PCM1 (E) granules around the centrosomes are shown. Average of three independent experiments are shown. *, P < 0.01; **, P < 0.05 compared with NS. (G) RPE1 cells were induced to quiescence and processed for immunofluorescence with anti-glutamylated tubulin and anti-PCM1 antibodies at indicated times after serum starvation. The percentages of RPE1 cells with primary cilia or accumulation of PCM1 granules around centrosomes were determined. Average of two independent experiments is shown.