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Fig. 3 | Molecular Neurodegeneration

Fig. 3

From: Tetraspanin 6: a pivotal protein of the multiple vesicular body determining exosome release and lysosomal degradation of amyloid precursor protein fragments

Fig. 3

APP-CTF accumulates in endosomal compartments in TSPAN6-overexpressing cells. a Immunofluorescence microscopy of HEK293 cells overexpressing myc-tagged TSPAN6, fixed and probed with a rat monoclonal antibody against myc, a rabbit monoclonal antibody against APP-CTF (Y188), and mouse monoclonal antibodies against EEA1 and LAMP1 to visualize early and late endosomal compartments, respectively. Cells overexpressing myc-TSPAN6 show more APP co-localizing with EEA1 and LAMP1 (filled arrowheads) compared to non-overexpressing cells (empty arrowheads). b Quantification of the number of cells non-overexpressing or overexpressing TSPAN6 showing co-localization of APP-CTF with early (EEA1) or late endosomal (LAMP1) compartments. c Immunoisolation of LAMP1-containing compartments as described [33], after overexpressing Flag-tagged LAMP1 in non-overexpressing or overexpressing myc-tagged TSPAN6. The total inputs and the immunoisolated compartments were analyzed by western blot with specific antibodies against APP, APP-CTF, TSPAN6 and LAMP1. Statistical significance was determined by t-test (***p <0.001)

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