Fig. 2: Derivation of cortical neurons from iPSCs affected by methylmalonic aciduria.
From: Mitochondrial dysfunction drives a neuronal exhaustion phenotype in methylmalonic aciduria
a Schematic of the 2D neuron differentiation protocol used in this article. Created using BioRender Forny, P. (2025) https://BioRender.com/w70y299. b Staining for SSEA4 and SOX2 at day 0 for pluripotent stem cells (iPSC). Staining for Nestin (NES) and SOX2 at day 13 for neuroepithelium. Staining for PAX6 and SOX2 at day 21 for neural stem cells (NSCs). Staining for TUBB3 and EOMES at day 40 for cortical progenitors. Staining for TUBB3/TBR1 and MAP2/NeuN at day 50 signal postmitotic deep layer cortical neurons and pan-neuronal molecular markers in cultures, respectively. Scale: iPSCs and neurons have 10 µm bars. Neuroepithelium, NSC, and NPCs have 50 µm bars. c RT-qPCR of SOX2, PAX6, and EOMES in iPSCs at day 0, neuroepithelium day 13, and NSCs day 21. Datapoints are representative of at least 3 independent experiments and error is reported as standard deviation. Expression values are relative to measurements at day 0 for each cell line. d Immunocytochemistry of patient and control NSC and neurons. Scale, 10 μm. e Western blotting analysis of neurons using anti-MMUT. MMUT is anticipated at 83 kDa. The loading control ACTB is anticipated at 42 kDa. Uncropped membranes are available in Supplementary Fig. 12.
