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 Jun 12;8:8984. doi: 10.1038/s41598-018-27174-x

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

Microplate luminescent reader assay for ERK activity. (a) HCT116 cells expressing hyBRET-ERK were plated in 96-well plates and treated with serially diluted AZD6244, an MEK inhibitor. BRET was measured with a microplate luminescent plate reader. (b) Dose-dependent decrease of ERK activity by AZD6244 in HCT116 cells. Eight wells were used for each condition. The experiment was performed in triplicate. (c) Scheme of the multiplexed cell-based assay for ERK activity, cell viability and cell death. (d–f) PC9 cells expressing hyBRET-ERK were plated in 96-well plates and treated with serially diluted gefitinib, an EGFR inhibitor. ERK activity and live cells were quantitated by the BRET ratio (d) and the sum of cyan and yellow luminescence intensities (e), respectively. Dose-response curves of gefitinib for ERK activity (d) and live cells (e). Each dot is the average of two wells. Data were fitted by the Hill equation. (f) Correlation of ERK activity versus the number of live cells. For more details, see the Materials and Methods.