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

. 2020 Nov 24;6:62. doi: 10.1038/s41523-020-00204-6

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

© The Author(s) 2020

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

Fig. 2 — A Human and rat amino acid (AA) sequence of PTGS2 gene region used as immunogens (SP21 = black bold and CX229 = gray box). Potential posttranslational modification (larger font size) S-nitrosylation site is seen at cysteine 526 (red arrow), disulfide bond sites at AA 555 and 561, and a glycosylation site at AA 580. B COX2 immunoprecipitation (IP) of HCA-7 cell lysate using CX229 is recognized by CX229 (lane 1), but not SP21 (lane 2). On biochemical S-nitrosylation of the CX229 IP, SP21 regains reactivity to COX2 (lane 4). C Western blot analysis confirms recombinant human COX2 protein contains the S-nitrosylated form of COX2 as detected by a pan-nitrosylation specific monoclonal antibody (lane 1), as well as by assessment of nitrosylation modification using the Pierce S-nitrosylation kit and the anti-TMT antibody (lanes 2 and 3). D Western blot analysis confirms dose-escalating Na ascorbate treatment (−, +, ++) used for de-nitrosylation of recombinant human COX2 protein results in dose-dependent loss of SP21 signal (lanes 1–3). No or minimal loss of CX229 reactivity was observed after Na ascorbate treatment (lanes 4–6). Quantitation of COX2 western blots from three separate Na ascorbate experiments (right panel electrophoretograms).