Extended Data Fig. 3: Human tNeurons derived from sAD patients exhibit disease-related protein pathologies.
From: Proteostasis and lysosomal repair deficits in transdifferentiated neurons of Alzheimer’s disease
(a) IF images and quantification of proteostasis- and disease-associated protein markers, including p62/SQSTM1, ubiquitin, total Aβ and the toxic isoform Aβ42, hyperphosphorylated tau (pTau) and TDP-43 (pTDP-43) and nuclear-to-cytoplasmic (N-to-C) ratio of TDP-43 in tNeurons shown as full images for Fig. 1g. Cells are co-stained with Tuj1. White dash line represents the nuclear region (N). p62/SQSTM1: n = 62 (young), 62 (aged) and 66 (aged/sAD) cells; ubiquitin: n = 46 (young), 53 (aged) and 66 (aged/sAD) cells; total Aβ: n = 111 (young), 106 (aged) and 114 (aged/sAD) cells; Aβ42: n = 103 (young), 95 (aged) and 93 (aged/sAD) cells; pTau: n = 91 (young), 89 (aged) and 114 (aged/sAD) cells; pTDP-43: n = 91 (young), 74 (aged) and 95 (aged/sAD) cells; N-to-C ratio of TDP-43: n = 85 (young), 86 (aged) and 103 (aged/sAD) cells. Scale bar: 20 μm. (b) IF staining and quantification of small heat shock protein HspB1 in tNeurons. n = 80 (young), 111 (aged) and 134 (aged/sAD) cells. Scale bar: 20 μm. (c) Sandwich ELISA assay for detecting endogenous Aβ42 in total cell lysates of tNeurons. The values are revealed by a fold change relative to young tNeurons. n = 4 (young), 4 (aged) and 4 (aged/sAD) independent replicates. (d) IF staining of pTau and pTDP-43 with p62/SQSTM1 and ubiquitin in aged/sAD tNeurons (scale bar: 20 μm). Cells are co-stained with Tuj1 and DAPI. White dash line represents the nuclear region (N). Insert: higher magnification view of protein co-localization within individual neuron (scale bar: 5 μm). Arrowhead: protein co-localization. In panels a, b and c, the boxes show median and 1st and 3rd quartile and the whiskers extending 1.5 times the interquartile range from the boxes. Data show box-and-whisker plots of two to three independent experiments and cells from independent HC and AD patients. Statistical analysis is performed using One-Way ANOVA followed by Bonferroni post-hoc analysis. *P < 0.05, **P < 0.01 and ***P < 0.001. Source numerical data are provided.
