Fig. 5.

LUPIN release of cytotoxic duocarmycin analogue in MCF-7 cell culture. a Schematic representation of Duo-OMe (4) release by LUPIN with furimazine to generate DNA alkylating agent. b Quantification of cellular growth (MCF-7) by nuclei count. Bar graph of nuclei count across different furimazine and prodrug (PNA-Py²Duo 12) concentrations (left panel). Representative images of cells (right panel) across different prodrug (12) and furimazine concentrations imaged by fluorescence (Hoechst) and bright field (BF). The red square indicates the zoomed-in area below it. The bar graph is the average of three independent experiments ran in triplicates. Error bars show ±1 standard deviation from the mean. Statistics were calculated using a two-tailed t-test with unequal variances (Welch’s unpaired t-test). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar: 200 µM; zoom-in scale bar: 50 µM