Fig. 3.

Resolving expanded 4T1 cellular subpopulations post-RT. A FACS expression levels of Her2 and cMet following RT. B–D Correlation plots between Her2 and cMet (B), Her2 and EGFR (C), and MUC1 and cMet (D) in irradiated cells. E Correlation plot between Her2 and EGFR levels expressed in the cells found to harbor process 3 (only cells with significant amplitudes (λ₃(cell) values) were included in this plot, also see the “Methods” section). F Protein–protein networks were generated using single-cell SA analysis and STRING to assign the functional connections. To determine the direction of change in every protein (i.e., upregulation or downregulation) a sign of the amplitude in a process α in each cell was considered. Four unbalanced subnetworks (processes) out of 10 resolved in 4T1 (Additional file 3) are shown. G Each cell was assigned a barcode representing a cell-specific signature (CSSS). The most abundant (>1%) subpopulations are presented. H Based on these CSSSs the tumor was divided into distinct subpopulations. Quantification of subpopulations was performed using at least ~30,000 cells from each condition, which were obtained from at least three flasks and from at least three independent experiments for each time point. For A: statistically significant differences between control and 5 Gy; control and 15 Gy; and 5 Gy and 15 Gy were determined using a two-tailed Student’s t test (*P < 0.01)