Fig. 4: Apoptotic cell death in RTD MNs.
From: Caspase-dependent apoptosis in Riboflavin Transporter Deficiency iPSCs and derived motor neurons
A Confocal microscopy after double IF, using anti- Act casp-3 (in green) and anti-β III-Tubulin (in red) demonstrates remarkable immunoreactivity to the apoptotic marker in RTD cells. Higher magnification merge images (A’ column), clearly show localization of Act casp-3 in the cytoplasm of RTD cells and in smaller structures, identified as apoptotic bodies. A minimum of n = 3 and 9 images were quantized. Scale bar, 10 µm. B Confocal images after TUNEL assay show remarkably more numerous positive nuclei (in green) in RTD cells, as compared to healthy cells. All nuclei were labeled in blue by HOECHST. A minimum of 11 images and 2200 cells were counted. Scale bar, 50 µm. C, D Bar graphs showing the percentage of Act casp-3 and TUNEL positive cells, respectively. Data are presented as the mean ± SEM of three experiments (n = 3) and analyzed by Mann–Whitney test. ****p ≤ 0.0001; ***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05.
