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. 2012 Oct 3;7(10):e46546. doi: 10.1371/journal.pone.0046546

Figure 6. DH281, DH285 and DH287 induce spindle abnormalities and increased Wee1 protein levels in HeLa cells.

Figure 6

A. HeLa cells were incubated with DH281, DH285 and DH287 at 1.21 µM for 24 hr. After that, the cells were fixed and incubated with FITC-conjugated anti-α-tubulin antibodies (red). DAPI was used to stain nuclear DNA (blue). Cells were visualized and photographed using a confocal microscope with a 20× objective. B. The protein levels of Wee1 but not cyclin B1 increase after treatment with the beta-carboline derivatives. HeLa cells were treated with DMSO or DH281, DH285 and DH287 at 1.21 µM for 24 hr and then protein samples were prepared. The proteins levels of Wee1 and cyclin B1 were detected after Western blotting. GAPH is shown as a loading control. C. The quantitative analysis of Wee1 and cyclin B levels in HeLa cells treated with β-carboline compounds. HeLa cells were treated with the compounds as described in B and then incubated with anti-Wee1 or anti-cyclin B1 primary antibody and FITC-conjugated secondary antibody. FACS was then performed to measure the protein levels of Wee1 and cyclin B1. Here shows the average of relative protein levels from three separate experiments.