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Figure 1 | BMC Neuroscience

Figure 1

From: Squalestatin alters the intracellular trafficking of a neurotoxic prion peptide

Figure 1

MoPrP105-132 is found in lipid rafts. Neuroblastoma cells incubated for 30 minutes with 30 μM rhodamine-labeled MoPrP105-132 and Alexa Fluor 488 labelled CTxB for 30 minutes, the nuclei were revealed using Vectashield with DAPI (blue). (A) Images of MoPrP105-132 (red) and CTxB (green) staining were merged using OpenLab software to show co-localization (yellow). (B) Images of scrambled MoPrP105-132 (red) and CTxB (green) staining showed that there was no co-localization (yellow). (C) FRET analysis showing the raw data between rhodamine-labeled MoPrP105-132 and Alexa Fluor 488 labelled CTxB and (D), the proximity of the donor/acceptor reaction revealed by false-colour intensity. Images of neuroblastoma cells incubated for 30 minutes with 30 μM rhodamine-labeled MoPrP105-132 (red) and stained with FITC-labelled anti-caveolin-1 (green), showing co-localization (yellow). (E) FRET analysis showing the raw data between rhodamine-labeled MoPrP105-132 and FITC-labelled CTxB and (F), the proximity of the donor/acceptor reaction revealed by false-colour intensity. Scale bars, 5 μm. (H), Western-blot analysis of triton X-100 insoluble fractions (lipid rafts) and soluble fractions (non-raft) isolated from neuroblastoma cells incubated with 30 μM MoPrP105-132-FITC for 30 minutes, and probed for MoPrP105-132, caveolin-1, TfR and GM1 (CTxB).

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