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. 2022 May 18;23(10):5650. doi: 10.3390/ijms23105650

Figure 2.

Figure 2

TNF-α stimulation increases ROS levels to induce PINK1 accumulation in RASFs. (a) After pretreatment with NAC (5 mM) for 1 h, RASFs were stimulated with TNF-α (10 ng/mL) or CCCP (10 μM) for 24 h. Cells were stained with CM-DCFDA for 30 min. Immunofluorescence revealed that TNF-α induced ROS formation and NAC quenched ROS. Data are representative of at least three independent experiments. (b) Cells were stained with JC-10 for 30 min and subjected to flow cytometry. Data are representative of five independent RASF preparations. * p < 0.05; *** p < 0.001. (c) Western blot for PINK1 in RASFs stimulated with TNF-α (10 ng/mL) for 24 h + NAC (5 mM), revealing increased PINK1 levels with TNF-α stimulation and decreased PINK1 levels in TNF-α + NAC co-treated cells.