Fig. 6. A: Left image – A2780 cells incubated with the prodrug 2 (20 μM) and Mit-specific dye R123 (1 μM). Two channels were used for emission monitoring: Ch1 – λex = 365 nm, λem = 420–470 nm (for detecting PDFPs) and Ch2 – λex = 430–510 nm, λem = 475–575 nm (for detecting R123). Ch1 + Ch2 combination is provided. Middle image – A2780 cells incubated with the prodrug 2 (20 μM) and nuclei-labelling NUCLEAR-ID® red dye (NIRD). Two channels were used for emission monitoring: Ch1 and Ch3 – λex = 538–562 nm, λem = 570–640 nm (for NIRD). Ch1 + Ch3 combination is shown. Right image: The same as the middle image, except that N-acetyl cysteine (NAC, 2 mM) was added. Zoomed-in (×2.5) images are shown in white boxes. B: Cell cycle distribution of A2780 cells treated for 72 h either with DMSO (1%, v/v, carrier) or HO-cpt (5 nM) or the prodrug 2 (0.5 μM). C: Quantification of the data shown in B. G0G1/(S + G2) – the number of cells in phases G0 and G1 divided by the number of cells in phases S and G2. D: The prodrug 2 (20 μM) and NIRD incubated either with human neutrophils (left image) or human neutrophils pre-incubated with PMA (right image). Combined “Ch1 + Ch2” images are shown (Ch1 and Ch2 are defined in in inset A). Right plot – % dead cells in human neutrophils incubated with the carrier, the prodrug 2 and HO-cpt (both 20 μM) for 2 h. E: Monitoring the fluorescence characteristic for PDFPs in immature cells (lineageNEG Sca-1+ c-kit+ (LSK) and myeloid progenitor (MP) cells) from BM in mice injected i.p. on days 0, 2 and 4 with the prodrug (12 mg kg−1). F: Monitoring of the number of LSK and MP cells in BM of mice treated as described in E. The analysis was conducted on day 7. Paired t-test was conducted between days 0 and 7. *: P < 0.05; **: P < 0.01; ***: P < 0.001.
