Fig. 3

Catalase-deficient mature adult mice induce leaky lysosomes. A Protein was extracted from liver lysates from WT and catalase-KO mice at 9 W and 53 W.Immunoblot analysis was performed using whole-cell lysates with LAMP1 antibody. B Quantified protein level of LAMP1 normalized with actin. C Representative fluorescence images of MEFs fixed and immunostained with anti-LAMP1 (red) and DAPI (blue). Scale bar represents 20 μm. D Intensities of lysosomal area showing red fluorescence (corresponding to control) and fluorescence intensity of MEFs. E Representative fluorescence images of lysotracker for WT and catalase-KO MEFs at P2 and P5 levels. Scale bar represents 20 μm. F Liver lysates from WT and catalase-KO mice at 9 W and 53 W were fractioned as described in the Materials and Methods. Equivalent volumes of each fraction were subjected to immunoblotting using the indicated antibodies. G–I Quantified protein level of cathD, cathB and LAMP1 on pellet and supernatant fractionations, normalized with actin. J Relative cath D levels were measured from liver lysates of mice, as indicated in A. Bar graph represents mean ± SD (n = 3 experiments). *P < 0.05; **P < 0.0001; ns: non-significant