Fig. 1.

Centrinone and CFI-400945 have different effects on NCI-H1299 lung cancer cells. (A) NCI-H1299 cells (American Type Culture Collection; grown on plastic in RPMI medium with 10% FBS) were stained for DNA (cyan), γ-tubulin (green), and Cep192 (red) after treatment with DMSO [0.02% (vol/vol)], centrinone (150 nM), or CFI-400945 (50 or 500 nM) for 7 d. Cells were replated on poly-d-lysine–coated glass coverslips, processed for immunofluorescence (3), and imaged on a DeltaVision Elite (GE Healthcare Life Sciences) equipped with a scientific complementary metal-oxide–semiconductor camera (pco.edge 5.5; PCO) using a 60× 1.42 N.A. PlanApo N or a 40× 1.35 N.A. UApo objective (Olympus). Lower magnification merged panels (Left) and magnified single-channel images of the regions in the yellow dashed boxes (Right) are shown. (Scale bars: Left, 20 μm; Right, 5 µm.) (B and C) Quantification of percent multinucleation (B) and centrosome number (γ-tubulin and Cep192-colabeled foci) (C). This manual analysis does not consider the extent of multinucleation. CFI-400945–treated cells are massively multinucleated; centrinone-treated cells are not.