Fig 1.
OdDHL has a cell-specific and significant impact on MDA-MB-231 viability in all culture conditions.
Significant differences in mean MDA-MB-231 viability at 100, 200 and 400 μM concentrations relative to the control in hypoxia and normoxia in 2D (A) and 3D and 2D in normoxia (B); *** = p-value < 0.0001, ** = p-value < 0.01, * = p-value < 0.05 based on Tukey post-hoc differences. Significant differences in MDA-MB-231 viability relative to the control between culture conditions with 400 μM treatment in hypoxia and normoxia in 2D (C) and 3D and 2D in normoxia (D); *** = p-value < 0.0001, ** = p-value < 0.01 based on least mean contrasts. (E) Photomicrographs of MDA-MB-231 cells cultured in normoxia (2D), hypoxia (2D), and 3D (normoxia) with fluorescence microscopy (with DAPI staining; scale bar = 100 μm) at 100X magnification after 24 hours of OdDHL treatment. (F) Photomicrographs of MCF-10A cells cultured in normoxia (2D), hypoxia (2D), and 3D (normoxia) with fluorescence microscopy (with DAPI staining; scale bar = 100 μm) at 100X magnification after 24 hours of OdDHL treatment. Differences in MCF-10A viability relative to the control at 100, 200 and 400 μM concentrations in hypoxia and normoxia in 2D (G) and 3D and 2D in normoxia (H). Error bars represent standard error of the mean.
Fig 2.
OdDHL has a cell-specific and differential impact on MCF-DCIS.com cells dependent on culture condition.
Significant differences in mean MCF-DCIS.com viability relative to the control at 100, 200 and 400 μM OdDHL concentrations in hypoxia and normoxia in 2D (A) and 3D and 2D in normoxia (B); *** = p-value < 0.0001, ** = p-value < 0.01, * = p-value < 0.05 based on Tukey post-hoc differences. Significant differences in MCF-DCIS.com viability relative to the control between culture conditions with 400 μM OdDHL treatment in hypoxia and normoxia in 2D (C) and 3D and 2D in normoxia (D); *** = p-value < 0.0001 based on least mean contrasts. Significant differences in mean MCF-10A, MCF-DCIS.com, MDA-MB-231 viabilities relative to the control at 400 μM OdDHL concentrations in hypoxia and normoxia in 2D (E) and 3D and 2D in normoxia (F); *** = p-value < 0.0001 based on Tukey post-hoc differences. Error bars represent standard error of the mean.
Fig 3.
400 μM OdDHL treatment significantly impacts the proliferation rate of MDA-MB-231 and MCF-DCIS.com cells in all conditions.
Significant differences in the mean percent of MDA-MB-231 cells entering S-phase of proliferation in the 0.6% DMSO and 400 μM OdDHL treatment groups in hypoxia and normoxia in 2D for N = 4 (A) and 3D and 2D in normoxia for N = 3 (B); *** = p-value < 0.0001 based on least mean contrasts. Significant differences in the mean percent of MCF-DCIS.com cells entering S-phase of proliferation in the 0.6% DMSO and 400 μM OdDHL treatment groups in hypoxia and normoxia in 2D for N = 3 (C) and 3D and 2D in normoxia for N = 3 (D); ** = p-value < 0.001, * = p-value < 0.05 based on least mean contrasts. Error bars represent standard error of the mean.
Fig 4.
400 μM OdDHL treatment significantly increases necrosis of MDA-MB-231 cells in all conditions.
(A) Differences in the annexin V-FITC staining mean gray value (MGV) for MDA-MB-231 cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3). (B) Differences in the propidium iodide staining MGV for MDA-MB-231 cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3); ** = p-value < 0.01, * = p-value < 0.05 based on least mean contrasts. (C) Differences in the annexin V-FITC staining MGV for MCF-DCIS.com cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia (N = 3), 2D/hypoxia (N = 4), and 2D/normoxia culture conditions (N = 4); *** = p-value < 0.001 based on least mean contrasts. (D) Differences in the propidium iodide staining MGV for MCF-DCIS.com cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3); ** = p-value < 0.01, * = p-value < 0.05 based on least mean contrasts. (E) Differences in the annexin V-FITC staining MGV for MCF-10A cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3); ** = p-value < 0.01 based on least mean contrasts. (F) Differences in the propidium iodide staining MGV for MCF-10A cells treated with 400 μM OdDHL, normalized to 0.6% DMSO control for 3D/normoxia, 2D/hypoxia, and 2D/normoxia culture conditions (N = 3); ** = p-value < 0.01 based on least mean contrasts. Error bars represent standard error of the mean.