Fig. 5: CatB deficiency promotes PEG3 degradation by impairing autophagy-lysosome activity and enhancing ubiquitin-proteasome function.
From: Cathepsin B deficiency disrupts cortical development via PEG3, leading to depression-like behavior
A Relative mRNA expression of PEG in the cortex of E12.5. WT (n = 4) and CatB−/− (n = 3) mice. B Immunoblot analysis of PEG3 in the N2a cells without or with 10 μM CA-074Me for 12 h. C Quantification of PEG3 in the immunoblots shown in (B). E Immunoblot analysis of PEG3 in the N2a cells exposed or not to 100 μg/ml CHX, 10 μM CA-074Me or a combination of both for 0, 1, 6, and 24 h. E Quantification of PEG3 in the immunoblots shown in (D). F Immunoblot analysis of LC3B in the N2a cells exposed or not to 10 μM CA-074Me. G Quantification the ratio of LC3B II/LC3B I in the immunoblots shown in (F). H Confocal image of the mCherry-GFP -LC3B signal in N2a cells exposed or not to 10 μM CA-074Me. The scale bar was 15 μm. I Quantification of the GFP-mCherry+ (red dots) and GFP+mCherry+ (yellow dots) positive LC3B puncta in (H) (n = 4 cells per group, the data were obtained from three independent experiments.). J Transmission electron microscope images of the autophagosome (red arrow) and autolysosome (yellow arrow) in N2a cells exposed or not to 10 μM CA-074Me. Scale bars were 1 μm and 500 nm. K, L Quantification of autophagosome (K) and autolysosome (L) shown in (H) (n = 9 cells per group). M Immunoblot analysis of ubiquitin in the N2a cells exposed or not to 10 μM CA-074Me. G Quantification of ubiquitin in the immunoblots shown in (M) (n = 3 per group, the data were obtained from three independent experiments. O Immunoblot analysis of PEG3 in the N2a cells exposed or not to 100 nM MG132, 10 μM CA-074Me or a combination of both for 12 h. P Quantification of PEG3 in the immunoblots shown in (O). (n = 3 per group, the data were obtained from three independent experiments.) *P < 0.05, ***P < 0.01, ***P < 0.001. Error bars represent mean ± SEM.
