Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Jul 3;8:10004. doi: 10.1038/s41598-018-28296-y

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Blocking formation of active autophagosomes with PI3K inhibitor LY294002 and Atg5 siRNA leads to failure of clearance of prion aggregates. SMB.s15 cells were treated with 40 μM PI3K inhibitor LY294002 for 16 hrs to block the formation of autophagosomes. (a) Representative immunoblot to show reduced LC3-I and LC3-II after LY294002 treatment, lane 3 and 4. (b) Representative TEM image showing aggregates scattered in the cytoplasm with empty vesicles after treatment with 5 μM spermine for 72 hrs, LY294002 and 10 μM CQ for the last 16 hrs. (c). Representative immunofluorescence images showing reduced colocalisation of LC3 with PrP^Sc aggregates after treatment with 5 μM spermine for 72 hrs, LY294002 and 10 μM CQ for the last 16 hrs. (d) Significantly reduced colocalisation between LC3 and PrP^Sc aggregates on LY294002 treatment. For the Atg5 siRNA treatment, the cells with either treated with 10 μM of CQ or 5 μM spermine and10μM of CQ for 24 hrs post transfection. (e) Representative immunofluorescence images showing reduced LC3 punctae and increased prion aggregates on treatment with Atg5 siRNA when compared to scrambled control siRNA. Treatment with spermine does not reduce the number of aggregates after Atg5 siRNA treatment (last lane). (f) Western blot to show reduced Atg5 expression on treatment with Atg5 siRNA when compared to scrambled control siRNA and the loading control β- actin. (g) Graph showing a significant increase in the number of PrP aggregates on spermine treatment post Atg5 siRNA treatment compared to spermine treated cells from scramble control siRNA. N = 2, Scale bar = 10 μm. Respective full length blots are shown in Supplementary Fig. 3. Abbreviations used: Chloroquine (CQ), Transmission Electron Microscopy (TEM).