Fig 1: Accumulative cytotoxic effect of doxorubicin and tumor cells on HUVECs. Representative fluorescent images of actin filaments (phalloidin, green fluorescence) showing morphology of endothelial cells stimulated by different concentrations of doxorubicin as well as MCF7 cells for 24 h at low (A) and high magnifications (B). Representative immunofluorescent images of ICAM1 (red fluorescence) of HUVECs stimulated by different concentrations of doxorubicin as well as MCF7 cells for 24 h at low (C) and high magnifications (D). Representative immunofluorescent images of THP1 (orange fluorescence) and endothelial cell (green fluorescence) adhesion stimulated by different concentrations of doxorubicin in the presence of MCF7 cells at high magnifications (E). Quantitation of cell numbers per view is measured and presented in panel (F). Quantitation of mean fluorescence intensity is measured and presented in panel (G). Corresponding quantitation of THP1 cell adhesion is shown in panel (H). Levels of eNOS (I) and ET-1 (J) in the culture medium were measured. Bars represent 50 and 100 µm. EC: endothelial cell; Dox: doxorubicin. Statistical analysis was performed by one-way ANOVA. n = 5 per group. *P < 0.05, **P < 0.001, ***P < 0.001, and ****P < 0.0001.
Fig 2: Cytotoxic effect of doxorubicin on HUVECs. Representative fluorescent images of actin filaments (phalloidin, green fluorescence) showing morphology of endothelial cells stimulated by different concentrations of doxorubicin for 24 h at low (A) and high magnifications (B). Representative immunofluorescent images of ICAM1 (red fluorescence) of endothelial cells stimulated by different concentrations of doxorubicin for 24 h at low (C) and high magnifications (D). Representative immunofluorescent images of THP1 cell (orange fluorescence) and endothelial cell adhesion (green fluorescence) stimulated by different concentrations of doxorubicin at high magnifications (E). Quantitation of cell numbers per view is measured and presented in panel (F). Quantitation of mean fluorescence intensity is measured and presented in panel (G). Quantitation of THP1 cell adhesion is shown in panel (H). Levels of eNOS (I) and ET-1 (J) in the culture medium were measured. Bars represent 50 and 100 µm. n = 5 per group. EC: endothelial cell; Dox: doxorubicin. Statistical analysis was performed by one-way ANOVA. n = 5 per group. *P < 0.05, ***P < 0.001, and ****P < 0.0001.
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