Fig. 2: Clonal FAD hNPCs produce high levels of Aβ with different Aβ42/40 ratio.

a Representative confocal images of ReN-mGAP (mixed) and ReN-mGAP10#D4 (clonal) cells. 2D-cultured cells were expanded on glass-bottom dishes. Cells were imaged with GFP and mCherry which represent APPSL and PS1ΔE9, respectively, using confocal microscopy. Scale bars represent 200 μm. b Expression of APP and PS1ΔE9 in clonal hNPCs. Cells were prepared in 6-well plates and grown for 24 h. Expression levels of APP and PS1ΔE9 were monitored by Western blot analysis. Levels of nicastrin and PS1-CTF were also monitored. β-Actin was used as a loading control. Asterisk represents a nonspecific band. c Comparison of Aβ38, Aβ40 and Aβ42 levels between mixed and clonal hNPC lines. Indicated mixed and clonal cells (3 × 10⁶) were grown in Matrigel-coated 6-well plates for 24 h. After changing media with 1 ml of fresh expansion media, cells were incubated for an additional 24 h. Conditioned media was analyzed by MSD Aβ assay to measure concentrations of secreted Aβ species. d Aβ42/40 ratios were determined. All values were combined to express as mean ± SEM of three independent repeats (black dots). Statistical significances were determined by one-way ANOVA with Tukey’s multiple comparisons test.