Fig. 5.

a The number of MIC-A/-B⁺ cells increased in drug-resistant MCF7. White peaks represent ESA⁺ cells. Black peaks represent the MIC-A/B⁺ CD44⁺ CD24^lo cells. b–e Co-culture of GEM^Res MCF7 cells with Notch-1 peptide-activated PBMC decrease the NICD-Notch⁺ cell numbers. Surviving NICD⁺ MCF7 cells after co-culture with: b no effectors. c IL-2 activated PMBC. d Notch-1 peptide-activated PBMC. e Numb-1 peptide-activated PMBC. The % of NICD⁺ cells is shown in the upper right quadrant. The decrease in NICD⁺ cells in relation to the NICD⁺ cells in panel (b) was: IL-2 activated PBMC, 4.4%; Notch-1-activated PBMC, 68.5%; and Numb-1-activated PBMC, 25.3%. Note: the amount of free NICD in GEM^Res MCF7 cells was lower than in UT-MCF7 cells (Fig. 3). f Numb-1 peptide-activated PBMC eliminate CD44⁺ cells from UT-MCF7 cells. 50,000 GEM^Res MCF7 cells were cultured with indicated effectors for 5 days. Then live cells were collected and analyzed for expression of CD44 and CD24 in the same experiment. Analysis was performed in large tumor cells of similar cellularity (FS: 800–1000, side scatter 100–800). All indicate all live cells regardless of phenotype, CD44^hiCD24^lo indicate CSC-like cells, CD44^hiCD24^hi indicate potentially metastatic MCF7 cells, CD44^loCD24^lo indicate non-invasive MCF7 cells. Open columns no effectors, NP, dotted columns MCF7 cultured with IL-2- activated PBMC, NO, dashed columns left to right MCF7 cultured with Notch-1-activated PBMC, NU, textured columns MCF7 cultured with Numb-1- activated PBMC