Interphase centriole centrin-2 puncta are marginally increased by CHC17 and CLC depletion. (A) HeLa–centrin-2–GFP cells were treated (72 h) with siRNA, as indicated (top). Depletion of proteins (left) was detected by immunoblotting siRNA-treated cell lysate. Molecular mass marker position is shown on the right. (B) HeLa–centrin-2–GFP cells were treated (72 h) with siRNA, as indicated (left), and then processed for IF to detect PCNT2 and CHC17. Centrin-2–GFP was visualized in green. Red-green merge is yellow. Bars, 10 µm. (insets) Boxes in each frame are magnified threefold. 2D confocal images are shown with the corresponding brightfield image. Bars, 2 µm. (C) The percentage of HeLa–centrin-2–GFP cells with more than four centrin-2 puncta per cell in interphase (representing G1, S, and G2 phases) was quantified for ≥50 cells for each siRNA treatment indicated. Centrin-2 puncta were identified by centrin-2–GFP that colocalized with PCNT2 detected by IF. Examples of interphase cells with more than two centrin-2 puncta are seen in B for the indicated siRNA treatments. Data represent the mean ± SEM for four independent experiments.