Figure 3: Inhibition of TBK1 affects mitosis and enhances 4n+ population.

(a) Cell cycle analysis of asynchronously growing H460 cells in the presence or absence of BX795 by flow cytometry. (b,c) Histograms summarizing cell cycle analysis of control and BX795-treated asynchronously growing H460 (b) and AALE cells (c). (d,e) Cell cycle analysis combined with pH3S10 staining of synchronized H1650 cells released for 9 h in the presence of BX795 by flow cytometry. (f) Cell cycle analysis of H1650 shcontrol cells (transduced with lentivirus containing scrambled sequence) or H1650 shTBK1 cells (transduced with lentivirus containing shRNA targeting TBK1) synchronized by double-thymidine block and released for 9 h by flow cytometry. In panels a, d and e, the y axis shows the quantification of cells stained for H3pS10 and the x axis shows DNA content (DAPI staining). (g) Graphical representation of the flow cytometry data showing reductions in the number of phospho histone H3 positive/mitotic cells when TBK1 is depleted in H1650 and H460 cells. (*P<0.05, t-test) (h) Graphs indicating the inhibition of cells undergoing mitosis when TBK1 was depleted in asynchronously growing H1650 and H460 cells. (*P<0.05; ^#P<0.067; t-test). Error bars represent s.d. Two replicates are included in these experiments.