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. 2016 May 20;20(9):1686–1695. doi: 10.1111/jcmm.12863

Figure 5.

Figure 5

Mfn2 knockdown leads to impaired γ‐secretase activity and maturation. (A) Protein expression of APP, BACE1 and γ‐secretase components was assessed by Western blot analysis of cell homogenates from NC and Mfn2 siRNA‐treated cells. (B) γ‐secretase activity assay using a membrane fraction from cells treated with NC or Mfn2 siRNA in the presence or absence of γ‐secretase inhibitor L‐685,458. APP fragments (C99, C83, AICD) were detected using the Y188 antibody. (C) Intact cells were treated with NC or Mfn2 siRNA and incubated with specific neprilysin peptide. Neprilysin activity was investigated by measuring the fluorescence of cleaved peptide (arbitrary units). Western blots of cell homogenates were performed. n = 4 (independent experiments), with duplicates or triplicates for each condition. Results are shown in mean ± S.E.M. Mann–Whitney U‐test was used for statistical analysis NC: negative control; Mfn2 siRNA: siRNA for Mfn2 mRNA.