Figure 4. A pathway analysis of the proteomic response to (A) hydroxyurea and (B) RO-3306 treatment.
Boxes containing large arrows and circles show the enriched KEGG and REACTOME pathways in each treatment. The direction of the arrows and colour indicates whether the pathway is up- (red) or down- (blue) regulated compared to asynchronous cells. The green lines and their thicknesses indicate the overlap between pathways. Proteins in individual pathways are shown as rounded squares and are connected by grey lines where protein–protein interactions have been reported. The colouring indicates the direction of the fold change and the shading represents the magnitude. (C) Immunofluorescence flow cytometry of asynchronous cells stained with αCDT1 antibody (AF-488 secondary conjugate, y-axis) and a DNA-binding dye (DAPI, x-axis). CDT1 protein expression is high in G1 cells, low in S-phase cells, and intermediate in G2&M cells. Comparison of CDT1 expression (D) and the relative ratio of CDT1:Geminin expression (E) measured by immunofluorescence flow cytometry in mock (red line) vs RO-3306 treated (blue line) G2&M cells gated by DNA content.