Extended Data Fig. 2: MEF2C-WT, but not MEF2C-L35P, could rescue impaired neuronal dendritic and axonal morphological development in Mef2c knockdown neurons.
From: Whole-brain in vivo base editing reverses behavioral changes in Mef2c-mutant mice
(a) Representative image of immunofluorescent staining for GFP (green), MAP2 (red), and DAPI (blue) of cultured E14.5 mouse cortical neurons. Mouse cortical neurons were transfected with DsRed shRNA (Ctrl, n = 39), shMef2c (n = 36), shMef2c with human MEF2C-WT (n = 40) or MEF2C-L35P (n = 32) respectively at DIV1 via Lipofectamine 3000. Neuronal cells were immunofluorescent stained at DIV10. Scale bars, 60μm. Quantitative analysis of neuronal total dendritic length (b) and branch numbers (c) of neurons transfected with Ctrl, shMef2c, shMef2c + MEF2C-WT, and shMef2c + MEF2C-L35P. (b: all P < 0.0001; c: shMef2c+WT versus shMef2c + L35P: P = 0.0002) (d) Representative immunofluorescent staining images for GFP (green), MAP2 (red), and DAPI (blue) of cultured E14.5 mouse cortical neurons. Neuronal cells were transfected with DsRed shRNA (Ctrl, n = 34), shMef2c (n = 37), shMef2c with human MEF2C-WT (n = 33) or MEF2C-L35P (n = 34) respectively at DIV1. Neuronal cells were immunofluorescent stained at DIV4. Scale bars, 60μm. Quantitative analysis of neuronal axonal (e) and total neurite length (f) of neurons transfected with Ctrl, shMef2c, shMef2c + MEF2C-WT, and shMef2c + MEF2C-L35P. n is total neuron numbers collected from more than 3 independent experiments. (e, f: all P < 0.0001) Statistical values represent the mean ± s.e.m. ***P < 0.001, ****P < 0.0001, (b, c, e, f) One-way ANOVA.
