Fig. 3

Neurons obtained from sAD-hiPSCs recapitulate the typical hallmarks of AD. A Immunofluorescence staining for Aβ using 6E10 antibody (red), DAPI (nuclei, blue), and EGFP (neurons, green) in HS and sAD hiPSC-derived neurons. sAD neurons show strong 6E10 staining, indicating the presence of Aβ pathology. Scale bars: 50 µm. B Quantification of 6E10 median fluorescence intensity (in arbitrary units, a.u.) showing a significant increase in Aβ accumulation in sAD-hNs neurons compared to HS-hNs; (n = 5/group). C Representative dot blot images of Aβ levels in DIV30 HS- and AD-hNs lysates; (D) Bar graphs showing the band intensity analysis of Aβ in HS- and sAD-hNs. Red Ponceau, RP, was used as loading index and used to normalize sample; synthetic oligomeric Aβ42 (200 nM) was used as positive control; (E) Representative WB images of p-TAU (Thr181, Ser199, Thr205, Thr217 and Thr231) levels in DIV30 HS- and sAD-hNs lysates; (F) Densitometric analysis of p-TAU (Thr181, Ser199, Thr205, Thr217 and Thr231), normalized to total human TAU (Ht7) protein levels; n = 5/group. GAPDH was used as a loading control;; norm. normalized protein levels. Each dot represents the average of three independent experiments conducted on n = 5/group. Data are presented as mean ± SEM, n.s. p > 0.05, ** p < 0.01 vs HS-hNs, assessed by Mann Whitney Rank Sum test