Fig. 2: Evaluation of hAMSC oxidative stress after 24 h of exposure to increasing BPA concentrations. | Cell Death Discovery

Fig. 2: Evaluation of hAMSC oxidative stress after 24 h of exposure to increasing BPA concentrations.

From: Bisphenol-A disrupts mitochondrial functionality leading to senescence and apoptosis in human amniotic mesenchymal stromal cells

Fig. 2

MtROS production in hAMSC was quantified in flow cytometry and in immunofluorescence using the MitoSOX Red fluorescent dye, after 3 and 24 h of exposure to increasing concentrations of BPA (0.05, 0.1, 0.2, 0.3, 0.35, and 0.4 μM). Results acquired in flow cytometry are presented as the percentage of MitoSOX Red-positive cells (A). Mitochondrial membrane potential (Δψm) variation was assessed using the JC-1 probe and representing the ratio, indicative of mitochondrial depolarization, following BPA exposure (B). Immunofluorescence images of hAMSC after 24 h of BPA exposure show MitoSOX Red-positive cells (red signal) overlaid on bright-field images (C), acquired at ×20 magnification (scale bar, 50 μm). The total number of MitoSOX Red-positive cells was quantified and is reported in (D). Fluorescence intensity of MitoSOX Red, measured as Normalized Integrated Density, is presented in (E). The antioxidant response of hAMSC was evaluated by quantifying Nrf2 and HO-1 gene expression after 24 h of BPA exposure, expressed as fold-change relative to the control condition (MetOH) (F). Histograms represent mean ± SD from n = 4 independent experiments. Statistical analysis was performed versus the control condition: p < 0.01 (*), p < 0.001 (**), p < 0.0001 (****).

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