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. 2019 Aug 15;9:11922. doi: 10.1038/s41598-019-47462-4

Figure 7.

Figure 7

Involvement of SNX33 in effects of donepezil on APP processing and Aβ content in culture media in primary cortical cells. (a–d) Effect of SNX33 knockdown on non-amyloidogenic APP processing in primary cortical cells. Cells were treated with 1 μM control or antisense MOs, in addition to 6 μL/mL of Endo Porter (EP) for 24 h, washed and treated with donepezil for 48 h. After donepezil treatment, cell lysate and culture media were subjected to western blotting. Representative images of western blotting analysis (a) and quantification of sAPPα/β-actin ratio (b) are shown. *P < 0.05; n.s., not significant. Data are expressed as mean ± SEM of n = 11–15 independent observations. (c,d) Representative images of western blotting analysis (c) and quantification of APP/β-actin ratio (d) are shown. n.s., not significant. Data are expressed as mean ± SEM of n = 16–18 independent observations. (e,f) Effect of SNX33 knockdown on donepezil-induced Aβ reduction. Cells were pretreated with MOs and EP for 24 h prior to treatment with 10 μM donepezil for 48 h. The amounts of Aβ40 (e) and Aβ42 (f) in culture media were measured by ELISA. **P < 0.01; ***P < 0.001; n.s., not significant. Data are expressed as mean ± SEM of n = 12 (e) or n = 13 (f) independent observations. SNX33, sorting nexin protein 33; MO, morpholino oligo; ELISA, enzyme-linked immunosorbent assay; APP, amyloid precursor protein.