Fig. 4

ER and the NFĸB pathway are active in MS Cluster 1 and required for MS formation. A IPA network analysis was performed for DEGs of the original MS Cluster 1. Two central nodes were identified, ESR1 (P-val = 4.73E-9) and NFĸB complex (P-val = 3.72E-3). Expression of ESR1 and ER target genes (B) and NFĸB family members and NFĸB target genes (D) are presented in dot plots, with color representing expression level and size representing the percentage of cells in each cluster expressing the gene. FEA was performed for ER (C) and NFĸB (E) gene signatures from MsigDB. AUC values are shown in a heatmap, and P-values are presented in Additional file 2: Supplemental Table 8. F Correlation between Hallmark ER or Hallmark NFĸB signatures with other Hallmark gene signatures in MS Cluster 1 was calculated using Pearson’s Correlation function in RStudio software. Correlation values are shown in a heatmap and P-values are presented in Additional file 2: Supplemental Table 9. G FEA was performed for the Hallmark TGFβ pathway gene signature on MCF-7 MS Clusters. Box plot shows signature scores per cluster and table indicates AUC and P-values for each cluster. H Multiple linear regression analysis between Hallmark ER, NFĸB and TGFβ pathway gene signatures was performed for MS Cluster 1. Single cells individually placed in 3D graph with each axis representing the z-score for each pathway and each rectangle representing individual cells