Fig. 3.

Aberrant intracellular Aβ accumulation in senescent primary neurons overexpressing mutant APP. a Schematic illustration of the experimental plan. Primary neuronal cultures were obtained from WT and G3Terc^−/− mice and infected at 7 days in vitro (DIV) with lentivirus expressing hAPP3xmut: mutated human APP (hAPP) carrying 3 AD-linked mutations (the Swedish [K670N/M671L], Florida [I716V], and London [V717I] mutations). Cell lysates were collected at 11 DIV. b Western blot analysis showing hAPP relative protein levels in WT and G3Terc^−/− neurons overexpressing APP3xmut. Actin was used as loading control, and the levels in the control group were set as 100%. Non-significant (two-tailed Student’s t-test, n = 8 cultures/group). The lack of hAPP detection in non-infected WT and G3Terc^−/− neurons is shown. c MSD Electro-Chemiluminescence Immuno-Assay (ECLIA) showing relative protein levels of human Aβ40 and Aβ42 in WT and G3Terc^−/− neurons overexpressing hAPP3xmut. Results were normalized by the amount of hAPP protein levels obtained in previous Western blot analyses, and the levels in the control group set as 100%. *P < 0.05 (two-tailed Student’s t-test or Mann–Whitney test, n = 8 cultures/group). hAβ signal was not detected in non-infected WT and G3Terc^−/− neurons. All data are presented as mean ± SEM