Fig. 4

Calcium deficiency induces selective autophagy in peroxisomes. A AML12 cells in calcium-deficient medium for indicated durations, immunostained with anti-PMP70 (red) and anti-LAMP1 (green), shown as representative fluorescence image. Scale bar represents 25 µm. B Quantification of PMP70 and LAMP1 co-localization. Data are expressed as means ± S.D. (n = 3, independent experiments, 30 cells were analyzed in each experiment), * p < 0.05. C Immunoblot analysis of AML12 cells in calcium-deficient medium for indicated durations. Whole cell lysates were reacted with anti-ATG5, anti-p62, anti-LC3II, and anti-β-actin. D Immunoblot analysis of ATG5 (+ / +) and ATG5 (-/-) MEF cells in calcium-deficient medium for 24 h. Whole cell lysates were reacted with anti-PMP70, anti-ATG5, anti-p62, anti-LC3II, and anti-β-actin. E Immunofluorescence of mRFP-GFP-SKL cells in calcium-deficient medium for indicated durations. Scale bar represents 25 µm. F Quantification of red fluorescence represents pexophagy. Data are expressed as means ± S.D. (n = 3, independent experiments, 30 cells were analyzed in each experiment), * p < 0.05. G RPE-1 cells in calcium deficient medium for indicated durations were fixed and immunostained with anti-PMP70 (red) and anti-NBR1 (green) shown as representative fluorescence image. Scale bar represents 25 µm