Figure 5.

Flow cytometry analysis to define the selectivity of the ODC_REMP and its activity to arrest cell cycle progression. (A) Pseudocolor blots demonstrating the difference in the size and granularity (forward- and side-scatter signal; FSC and SSC, respectively) of 300 untreated (CTRL) and ODC_REMP-treated 3Dcc^imm spheroids after dissociation into single-cell suspensions. (B) Propidium iodide staining was performed to detect the distribution of cells in the different stages of the mitotic cell cycle. (C) Expression of cell surface proteins shown through FACS analysis. The expression profile was captured after four and six repeated treatment administrations on day 5 (5 d) in comparison to the CTRL. Single and double protein expressions have been analyzed in comparison to cell surface proteins exclusively expressed on immune cells. The error is presented as the standard deviation. Significance was calculated for N = 3 independent experiments using a two-way ANOVA test; * p < 0.05, ** p < 0.01, *** p < 0.001. (D) Measurement of apoptosis through Annexin V staining demonstrating the increase in apoptotic cells after ODC_REMP treatment. (E) Comparison of the unstained control versus the untreated and ODC_REMP-treated 3Dcc^imm spheroids shown in histograms.