Fig. 6. The expression levels of mTORC1 and HDAC are higher in TNBC cell lines and TNBC patient tumors; co-inhibition of mTORC1, ESR1α, and HDACs reduces the viability of patient tumors’ fragments and CSCs.

a, b The expression of mTORC1 and HDAC genes in 10 TNBC cell lines and 55 TNBC patient samples were compared using the NCBI Gene Expression Omnibus (GEO2R). The GSE65216 samples were analyzed with the Affymetrix Human Genome U133 Plus 2.0 Array (GPL570). c Alamar blue viability analysis of three primary TNBC patient fragments (CRDCA, SEM-1, and ARI-1) and one patient-derived xenograft fragment (HCI-001). TNBC fragments were cultured and treated for 144 h with vehicle (DMSO, D), rapamycin (5 nM, R), tamoxifen (1 µM, T), valproic acid (250 µM, V), or VRT combination. d Representative flow cytometric data showing the percentages of CSC (CD44^high/+/CD24^low/−) subpopulation in patient-derived xenograft TNBC fragments after treatments as described in c. e Relative living CSCs (CD44^+/high/CD24^−/low and also negative for 7-AAD and Annexin-V) in TNBC patient tumor fragments after treatment as described in c