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

. 2019 Jan 23;9:410. doi: 10.1038/s41598-018-37207-0

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

© The Author(s) 2019

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

Pharmacologic inhibition of PERK signaling increases secretion of ^FTTTR^A25T in non-native conformations. (A) Illustration showing the three assays used to monitor total, tetrameric, and aggregate TTR in conditioned media. (B) Graph showing total (filled bars) and tetrameric (open bars) ^FTTTR^A25T in conditioned media prepared in the presence of Taf-S on HEK293T cells treated for 16 h with Tg (500 nM) and/or ISRIB (200 nM). Total and tetrameric ^FTTTR^A25T are normalized to vehicle. Relative populations of non-native TTR in each condition are shown. Error bars show SEM for n = 13 independent experiments. (C) Representative chromatogram showing ^FTTTR^A25T tetramers in conditioned media prepared in the presence of Taf-S on HEK293T cells treated for 16 h with Tg (500 nM) and/or ISRIB (200 nM). ^FTTTR^A25T tetramers were separated on anion exchange chromatography using the UPLC system and visualized by compound 1 fluorescence. (D) Graph showing normalized ^FTTTR^A25T tetramers in conditioned media prepared in the presence of Taf-S on HEK293T cells treated for 16 h with Tg (500 nM) and/or ISRIB (200 nM). Normalized TTR tetramers were calculated using the formula: normalized TTR tetramers for a given condition = (TTR tetramer signal condition/TTR tetramer signal veh)/(total TTR signal condition/TTR tetramer signal veh). Error bars show SEM for n = 13 independent experiments. (E) Representative Clear Native (CN)-PAGE immunoblot of ^FTTTR^A25T soluble aggregates in conditioned media prepared in the presence of Taf-S on HEK293T cells treated for 16 h with Tg (500 nM) and/or ISRIB (200 nM). Total ^FTTTR^A25T in each condition is shown in the SDS-PAGE immunoblot. (F) Graph showing the amounts of ^FTTTR^A25T soluble aggregates in conditioned media prepared in the presence of Taf-S on HEK293T cells treated for 16 h with Tg (500 nM) and/or ISRIB (200 nM). Aggregate ^FTTTR^A25T is normalized to vehicle. Error bars show SEM for n = 13 independent experiments. *Indicates p < 0.05, *** indicates p < 0.005 for a two-tailed paired t-test.