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

. 2017 Sep 13;26(24):4765–4777. doi: 10.1093/hmg/ddx350

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

© The Author 2017. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 7. — Poly PA inhibits poly GA toxicity in chick spinal cord: DPR versus RNA toxicity of G4C2 hexanucleotide repeats. (A) Cleaved PARP and TDP-43 were assessed in lysates, of HEK-293 cells co-transfected with GA (250 ng) and increasing concentrations of PA-EGFP (0-1000 ng). (B) PA-EGFP expression did not increase PARP cleavage. Increasing concentrations of PA-EGFP (0-1000 ng) were transfected without GA-EGFP. (C) High molecular weight GA-EGFP species is increased in a dose-response manner. Total cell lysates from GA-EGFP (50–1000 ng) transfected HEK-293 cells were used for anti-GA western blotting. (D) PA-EGFP inhibits GA-EGFP aggregation. The appearance of high molecular weight GA-EGFP species is inhibited by a gradual increase of PA-EGFP. (E) PA-EGFP rescues GA-EGFP- mediated toxicity in the chick spinal cord. Transverse sections of embryonic day 3.5 (E3.5) chick embryo spinal cords (dorsal at top), electroporated with DPR constructs as labelled – GA and PA embryos were electroporated with 2 µg of plasmid. GA-EGFP: PA-EGFP (2: 2) and GA-EGFP: PA-EGFP (3: 1) were electroporated with different concentrations (µg) of plasmids as indicated. Scale bar = 100 μm. (F) Quantitative analysis of TUNEL-positive cells showing that the number of TUNEL- positive GA-EGFP expressing cells are significantly decreased by PA-EGFP expression in chick spinal cord (***P<0.0001). (G) GA-EGFP and PA-EGFP expression in chick spinal cord were detected by immunostaining for GA and PA. Scale bar = 100 μm. DAPI and GFP show subcellular localization of DPRs by using DPR antibodies. Scale bar= 100 μm. (H) Comparison of DPR expression between synthetic DPR construct and G4C2 repeat construct by DPR immunostaining in chick spinal cord. The GA-EGFP, GP-EGFP, and GR-EGFP constructs (a, e, i) are transfected with the same amount of EGFP-G4C2x 72 repeat construct (c, g, k). DPR expression levels were compared with specific poly GA, poly GP and poly GR antibodies. Scale bar = 100 μm. The data were analysed from six embryos in three independent sets of experiments.