Figure 5.

FAM134B-mediated ER-phagy regulated AGEs-induced intracellular ROS accumulation, apoptosis, and senescence in human NP cells. After lentivirus and siRNA transfection were separately conducted for 72 and 48 h, the human NP cells were cultured with 200 μg/mL AGEs for 36 h. (a–d) FAM134B overexpression and knockdown efficacy were verified by western blot assay. (e–g) The intracellular ROS levels were probed using the fluorescent dye DCFH-DA and measured by flow cytometry. (h) Relative protein expression levels of the apoptotic and senescent associated proteins p53, p16, and cleaved caspase3 were evaluated using western blot assay. (i) Cell apoptosis was assessed by TUNEL staining; representative TUNEL immunofluorescent images and apoptotic ratio were quantitated, scale bar: 100 μm. (j) Representative SA-β-gal staining images and positive cell ratio were illustrated, scale bar: 50 μm. Data are represented as mean ± SD. ^∗∗P < 0.01, ^∗P < 0.05.