Fig. 2: Neuron migration and differentiation is inhibited by CatB inhibitor.
From: Cathepsin B deficiency disrupts cortical development via PEG3, leading to depression-like behavior
A Fluorescent images of Filamentous actin (F-actin, magenta) in N2a cells at 72 h after damage by scratch. The N2a cells were treated with 10 μM Pepstatin (aspartyl proteases inhibitor), 10 μM Z-FF-FMK (CatL inhibitor), 10 μM CA-074 (cell none-permeable CatB inhibitor) or 10 μM CA-074Me (cell-permeable CatB inhibitor) 1 h scratch. Scale bars were 5 μm. The right images are the close-ups of the boxes in the corresponding left images. B Quantification of the migration of N2a cells shown in (A). The gap width was quantitatively evaluated using Image J software. C Bright-field images of N2a and N3a cells at 72 h after damage by scratch. Scale bars were 50 μm. D Quantification of the migration of N2a and N3a cells shown in (C). E Fluorescent images of F-actin (green), MAP2 (magenta) with Hoechst (blue) in primary cortical neurons with CatB siRNA or CA-074Me treatment. Scale bars were 10 μm. F Immunofluorescent staining of Nestin (green) in NSCs at day 1, 3, 5, 8 and 11 after differentiation. Scale bars were 20 μm. G Immunofluorescent staining of MAP2 (green) with CatB (red) in neuron stem cells (NSCs) at day 1, 3, 5, 8, and 11 after differentiation. Scale bars were 20 μm. H Quantitative analysis of the fluorescent density of MAP2, Nestin and CatB shown in (E, F). I Immunofluorescent staining of MAP2 in WT NSCs, WT NSCs treated with CA-074Me, and NSCs of CatB−/− mice at day 5 and 8 after differentiation. J, K Quantitative analysis of the number and the mean branch number of MAP2-positive cells. All data were obtained from three independent experiments. **P < 0.01, ***P < 0.001. Error bars represent mean ± SEM.
