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. 2020 Jan 17;10:601. doi: 10.1038/s41598-020-57516-7

Figure 7.

Figure 7

BrainPhys increases mRNA levels of APP and APP-cleaving enzymes. NPCs differentiated in either BrainPhys medium or NMM for up to 35 days and the gene- and protein expressions of APP and the APP-cleaving enzymes α-secretase (ADAM10), β-secretase (BACE1) and γ-secretase (PSEN1) investigated using qPCR and western blot, respectively. (A) APP mRNA levels (I) show a significant increase in BrainPhys-cultured cells compared with NMM, while no change in intracellular full-length APP protein levels (II) is observed. Mean values of five separate experiments on neurons from two different iPSC lines (Ctrl1 marked with grey circles and ChiPSC22 marked with black circles) were analysed with Student’s t-test. Bars represent mean +/− SEM. ***p ≤ 0.001. One representative blot out of three is shown. (B) ADAM10 (a disintegrin and metalloproteinase domain-containing protein 10) mRNA levels (I) show a significant increase in BrainPhys-cultured cells compared with NMM, while no change in intracellular ADAM10 protein levels (II) is observed. BACE1 (beta-site APP cleaving enzyme 1) mRNA levels (III) show a significant increase in BrainPhys-cultured cells compared with NMM, while no change in intracellular BACE1 protein levels (IV) is observed. Both PSEN1 (presenilin1) mRNA levels (V) and protein levels (VI) increase significantly in the BrainPhys-cultured cells compared with NMM. Mean values of five (mRNA) or four (protein) separate experiments on neurons from two different iPSC lines (Ctrl1 marked with grey circles and ChiPSC22 marked with black circles) were analysed with Student’s t-test. Bars represent mean +/− SEM. **p ≤ 0.01, ***p ≤ 0.001. Full-length blots are presented in Supplementary Fig. 1.