Fig 1: Calcium deficiency-induced pexophagy is regulated by ROS-mediated TFEB activation. A Immunoblot analysis results of AML12 cells in calcium-deficient medium with or without NAC treatment. Whole cell lysates were reacted with anti-catalase, anti-Pex14, anti-PMP70, and anti-β-actin. B Immunofluorescence of mRFP-GFP-SKL cells in calcium-deficient medium for indicated durations with or without NAC treatment. Scale bar represents 25 µm. C Quantification of red fluorescence represent pexophagy. Data are expressed as means ± S.D. (n = 3, independent experiments, 30 cells were analyzed in each experiment), * p < 0.05. D AML12 cells in calcium-deficient medium with or without chloroquine treatment are immunoblotted. Whole cell lysates were reacted with anti-catalase, anti-Pex14, anti-PMP70, and anti-β-actin. E Cytosolic and nuclear fractions from AML12 cells with or without calcium for 24 h. Also includes treatment of NAC and chloroquine. Immunoblot the fractions with anti-TFEB. For the cytosolic and nuclear fractions, alpha-tubulin and CREB antibodies, respectively, were utilized as the controls