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. 2020 Jun 25;21(12):4516. doi: 10.3390/ijms21124516

Figure 3.

Figure 3

Stimulation of iPSC-derived neuronal cultures with the Aβ-S8C dimer. (A) Scheme illustrating the approach. Neurospheres were maintained for 4 months in culture, dissociated into single cells, differentiated for 6 weeks then stimulated with 500 nM of Aβ-S8C dimer for 72 h. Western blotting, microarrays and cytokine arrays were performed. (B-D) ELISA quantification of (B) total Aβ40, (C) total Aβ42 levels and (D) Aβ42/Aβ40+42 ratio from media collected from the interneuronal network and normalized to the total protein content. All data are presented as mean ± SEM from six independent experiments. (E) Representative Western blot images of endogenous TAU, phosphorylated TAU (Ser 202 and Thr 205), APP and the neural differentiation marker βIII-Tubulin after stimulation with 500 nM of the Aβ-S8C dimer. β-ACTIN was used as a loading control. (F-H) Quantification of (F) total TAU, (G) phosphorylated TAU and (H) APP levels. Results are normalized against β-ACTIN and shown as a percentage of control (CTR). All data are presented as mean ± SEM from 3 independent experiments. (I). Effect of the Aβ-S8C dimer on APP levels in iPSCs derived neuronal network (CON8, CON9, AD2-2 and AD2-4) compared to control. Data are presented as mean ± SEM from 3 independent experiments from 4 biological replicates. * p < 0.05, one-tail t-test versus control.