Fig. 6

Multilevel proteomics show differential apoptosis signaling. A Heatmap of KP and KPP phosphorylation sites after treatment with solvent control, KU-60019, radiation and combined treatment. Phosphosites (rows) and samples (columns) have been hierarchically clustered using Euclidean distance. Quantification values have been standardized using Z-scoring to account for different scales. Color scales indicate Z-scores. B Enrichment map showing Reactome pathways differentially regulated (log2 fold change differences > 0.5) between KP and KPP cells upon radiation. Related pathways are connected by edges. Node coloring corresponds to ReactomeFI functional enrichment score. All pathways shown are significantly enriched with an FDR < 0.05. C Heatmap showing total protein fold changes of apoptosis hallmark genes upon radiation and combinatorial treatment in KP and KPP cell lines. Clustering has been performed using hierarchical clustering with Euclidean distance. D Bar graph showing log2 fold changes for genes identified in cluster I from C. The data indicates that combinatorial treatment rescues the expression differences upon radiation between the two cell lines. E Annexin V/DAPI staining of KP6 and KPP4 cells with 3 h pre-treatment 3 µM KU-60019 and DMSO as control with and without irradiation 8 Gy, 96 h post irradiation. Supernatant of 96 h cultivation Medium was collected with trypsinized cells before staining. The lower right quadrant of the dot plots shows the apoptotic fraction measured with flow cytometer. The diagram shows the apoptotic fraction after 96 h with different treatments. Error bars: Standard deviation. Also see Additional file 1: Fig. S6