FAM134B-Mediated ER-phagy Upregulation Attenuates AGEs-Induced Apoptosis and Senescence in Human Nucleus Pulposus Cells

Figure 2

Intracellular ROS accumulation was involved in AGEs-induced senescence and apoptosis. The human NP cells were exposed to 200 μg/mL AGEs for different times (0, 6, 12, 24, and 36 h) or directly cocultured with 200 μg/mL AGEs and ROS inhibitor 10 μM NAC for 36 h. (a, b) The intracellular ROS levels were detected using the fluorescent probe DCFH-DA and measured by flow cytometry. (c, d) After labeled with DCFH-DA fluorescent probe, representative fluorescent images were acquired under a fluorescence microscope, scale bar: 100 μm. (g–j) Representative western blot bands of p16, p53, and cleaved caspase 3 and relative band density were quantified. (k, l) Representative images of immunofluorescence staining for p16 and cleaved caspase-3 in each group, with the relative fluorescence intensity quantified, scale bar: 50 μm. Data are represented as . , .

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