Fig. 5. Elevated p-tau levels in the PS1-E120K iPSC-derived neurons. (A~C) Western blot analysis showing a significant increase of phosphorylated tau (p-tau) including AT8, AT180 and p-Tau (Ser400, Thr403/Ser404) and total tau (t-tau, Tau5) proteins in the PS1-E120K iPSC-derived neurons compared with the control group. (D~F) Quantification of the western blots on the expression of AT8 (D), AT180 (E) and p-Tau (F) showing a significant increase of the expression of each in the PS1-E120K iPSC-derived neurons. (G) Immunocytochemical analysis showing the expression of AT8 (red) and MAP2 (green), counter-stained with DAPI (blue), in the PS1-E120K iPSC-derived neurons at 10 weeks of neuronal differentiation. Note that PS1-E120K iPSC-derived neurons exhibit a dramatic increase of phosphorylated-tau (AT8) both in the soma (arrow) and neurites (arrowhead) compared with the control group. Scale bar: 10 µm. (H~J) Quantification of the immunocytochemical analysis on the expression and localization of AT8 proteins with respect to MAP2-positive neurons. MAP2 expression was normalized against DAPI (G), whereas AT8 expression was normalized against MAP2 expression (I, J). Note that significant increase of AT8 expression both in the soma (I) and neurite (J) regions in the PS1-E120K iPSC-derived neurons. Representative western blot and ICC images were obtained from three independent experiments.
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