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 Mar 10;10:4429. doi: 10.1038/s41598-020-61370-y

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

N-glycosylation of hGPR176 and its naturally occurring nonsynonymous variants. (A) Reported rare nonsynonymous SNPs of hGPR176. Note that the S13G variant disrupts the N-X-S triplet sequence required for glycosylation at N11. (B) Immunoblot of Dox-treated Flp-In TREx293-hGPR176 (tet-on) cells with or without PNGase F treatment. Asterisk, nonspecific bands. (C) Immunoblots of Dox-treated (+) and untreated (−) Flp-In TREx293 (tet-on) cells for WT and N-ter mut (N4Q;N11Q;N17Q;N27Q) hGPR176. (D) GloSensor activity traces in Dox-treated (+) and untreated (−) Flp-In TREx293 WT and N-ter mut hGPR176 (tet-on) cells. Values are the means ± s.d. (n = 3 for each data point). (E) Relative protein expression levels of Dox-induced WT and N-ter mut hGPR176. Values are the means ± s.d. (n = 3 for each) of the relative band intensities in (C). ****P < 0.0001, two-tailed unpaired t-test. (F) Percent cAMP-inhibitory activity of N-ter mut hGPR176 relative to that of WT hGPR176. The AUC in (D) was used to evaluate the rate of cAMP inhibition. Data are the means ± s.d. of three replicates each. ***P < 0.001, two-tailed unpaired t-test. (G) SDS-PAGE/immunoblot profiles of naturally occurring hGPR176 variants. We generated Flp-In TREx293 tet-on stable cell lines expressing each variant with the constitutive expression of GloSensor. (H) Relative protein expression levels of WT hGPR176 and respective variants of hGPR176. Values are the means ± s.d. (n = 3 for each). *P < 0.05, versus WT, one-way ANOVA with Bonferroni post hoc test. (I) Percent cAMP-inhibitory activities of respective hGPR176 variants relative to that of WT hGPR176. Data are the means ± s.d. of three replicates each. **P < 0.001, *P < 0.01, versus WT, one-way ANOVA with Bonferroni post hoc test. (J) Relationship between protein expression level (x axis) and inhibition activity (y axis) of WT and respective variants of hGPR176. Values are the means ± s.d. (n = 3 for each data point). Correlation coefficient (r) and P value were calculated by Pearson product moment correlation coefficient analysis (r = 0.9511, P < 0.01).