Novel therapeutic diiminoquinone exhibits anticancer effects on human colorectal cancer cells in two-dimensional and three-dimensional in vitro models

Figure 9 Effect of diiminoquinone on established patient-derived organoids from colon cancer patient 3. A: Immunohistochemistry images of tissue derived from patient 3 stained with hematoxylin and eosin (HE). Immunofluorescent images of tissue stained with colon lineage epithelial markers cytokeratin (CK)19 and stem cell marker CD44. The nuclei were stained with anti-fade Fluorogel II with 4’, 6-diamidino-2-phenylindole (DAPI). Representative confocal microscopy images were acquired using a Zeiss LSM 710 laser scanning confocal microscope. Scale bar 20 μm; B: Immunofluorescent images of organoids derived from colon cancer patient 3 at generation (G)2 in the presence and absence of diiminoquinone (DIQ) treatment (0.5 and 1 μmol/L) stained with colon lineage epithelial markers CK19 and stem cell marker CD44. The nuclei were stained with anti-fade Fluorogel II with DAPI. Representative confocal microscopy images were acquired using a Zeiss LSM 710 laser scanning confocal microscope. Scale bar 20 μm. Representative bright-field images of organoids derived from colon cancer patient 3 at G2 in the presence and absence of DIQ treatment (0.5 and 1 μmol/L). Organoid formation count and size were calculated, and mean values were reported as mean ± standard error of the mean (^aP < 0.05, ^bP < 0.01, ^cP < 0.001). Images were visualized by Axiovert inverted microscope at × 5 and × 20 magnification. Organoid formation count (OFC) and size of G were calculated, and mean values were reported as mean ± standard error of the mean (^aP < 0.05, ^bP < 0.01, ^cP < 0.001). Scale bar 100 μm.