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Fig. 2 | BMC Molecular and Cell Biology

Fig. 2

From: Alix and Syntenin-1 direct amyloid precursor protein trafficking into extracellular vesicles

Fig. 2

Alix and Syntenin-1 drive mutant APP packaging into secreted vesicles. a Immunoblot analysis of cell lysates following activation of stably transduced inducible shRNA gene knockdowns targeting ESCRT proteins Hrs (shHrs) and Tsg101 (shTsg101), tetraspanin protein CD63 (shCD63), and Alix (shAlix) and Syntenin-1 (shSyntenin-1), and subsequent transfection of APPswe. Equal protein mass was loaded into gels. b Immunoblots of corresponding secreted EVs, with equal volume loaded. c) Comparison of endogenous APP secretion into EVs following Alix or Syntenin-1 knockdown. d) Immunoblot analysis of CTFβ peptides secreted into EVs. Quantitation of (e) APP and (f) CTFβ secreted into vesicles from shRNA-expressing cells, normalized to cellular levels. Blots are representative images from four to five repeated independent experiments. Statistical differences were determined by two-way student’s t-test. Nanoparticle tracking analysis showing (g) quantity and (h) size of small EVs from control, shAlix, and shSyntenin-1 cells with or without APPswe expression. One-way ANOVA was used to determine statistical significance across samples from three independent experiments. ***, p < 0.001; **, p < 0.01; *, p < 0.05

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