Figure 5. VDAC3 recruits Mps1 to centrosomes to regulate centriole assembly. (A and B) siCon or siVDAC3–1 (siVD3–1) RPE1 cells expressing GFP-PACT (green) or GFP-Mps1-PACT (green) were arrested in S-phase by a 24 h HU treatment and were examined for centriole numbers. (A) Shown are representative siVDAC3–1 cells stained for Cetn2 (red), γ-tub (magenta) and DNA (blue); GFP is green, bar = 5 μm. (B) Percentage of GFP-positive cells with ≥ 3 Cetn2 foci, values represent the mean ± SD for three independent experiments, 50–60 cells counted per replicate. (C–F) HeLa GFP-Mps1^∆12/13 cells transfected with control (siCon) and VDAC3–1 (siVD3–1) siRNA were arrested in S-phase by a 72 h HU treatment, with or without dox, to induce GFP-Mps1^∆12/13 [green in (C)], and stained for DNA (blue) and (C) γ-tub (red) or (D) CP110 (red) and Sas6 (green). Bar = 5 μm. (E) The percentage of cells with more than two centrosome equivalents (> 2 Sas6 foci or > 4 CP110 foci), values represent the mean ± SD for three independent experiments, at least 100 cells counted per replicate. (F) Immunoblots showing the expression of GFP-Mps1^∆12/13 (arrowhead), endogenous Mps1 (arrow) and depletion of VDAC3 (VDAC3a roughly 10%, VDAC3b roughly 80%), α-tub as loading control.