Fig. 1. Enrichment of CD44+CD24 cells within EAC cells using different concentrations of different chemotherapeutics . A. Dot plot of flow cytometry using FITC anti-CD44 and PE anti-CD24. Ehrlich cells were cultured in T-25 tissue culture flasks at a seeding density equals to 2 × 106 cells/flask. When the cells reached 80% confluency, various concentrations of CDDP, DOX, and PTX were added for 72 hrs. (n = 3 for each concentration). The media was then discarded and adherent cells were washed by PBS and detached by trypsin-EDTA for 5-10 min and the latter was then inactivated by FBS. Cells were then collected in tubes and analyzed by flow cytometry for CSCs markers.B. Bar chart of the percentages of CD44+CD24- cells of the various chemotherapeutics concentrations used showing the significance levels against control. CDDP at concentration = 50 µg/mL showed the highest enrichment of CSCs. One-way ANOVA was done. Data are presented as mean ± SD. ** p < 0.01, *** p < 0.001. EAC: Ehrlich ascites carcinoma, FITC: fluorescein isothiocyanate, CD: cluster of differentiation, PE: phycoerythrin, PBS: phosphate-buffered saline, FBS: fetal bovine serum, CSCs: cancer stem cells, CDDP: cisplatin, DOX: doxorubicin, PTX: paclitaxel.