Fig. 3
From: Deleterious mutations in ALDH1L2 suggest a novel cause for neuro-ichthyotic syndrome
Patient fibroblasts have lower ATP and energy index, and altered mitochondrial morphology in comparison with fibroblasts from parents or SLS patient. a Western blots assays of ALDH1L2 and ALDH3A2 in fibroblasts isolated from the patient (R), both parents (mother, M; father, F) and from an SLS patient (SLS). LR denotes patient’s fibroblasts transduced for ALDH1L2 expression. Numbers on the bottom of each panel indicate band intensity (arbitrary units) for ALDH1L2 and ALDH3A2 relative to the intensity of corresponding actin band. b Doubling time and energy index of different fibroblast cultures (cell labeling as in panel a). c ATP levels measured by a colorimetric assay in different fibroblast cultures. Three different samples (biological replicates) were used in this experiment; for each sample, 4 measurements (technical replicates) were performed and the average of these measurements were used to calculate mean ± SE. d TMRM (tetramethylrhodamine) to MitoTracker Green ratio in different fibroblasts. Six samples (biological replicates) were analyzed for each cell type. e Levels of ROS evaluated by confocal microscopy after DCF (2′,7′-dichlorodihydrofluorescein diacetate) staining in patient (R cells) and control (C cells) fibroblasts (values were calculated from the analysis of 10 cells for each cell type; laser power was kept uniform for all measurements). f Confocal images (108×) of different fibroblast cultures (as in panel a). Live cells were stained with Hoechst (nucleus staining, light-blue), MitoTracker Green (mitochondrial staining, green), or TMRM (mitochondrial staining, red); scale bars, 10 µM. For panels c, d, *p < 0.05; **p < 0.01; ***p < 0.001
