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

. 2022 Mar 28;23(7):3702. doi: 10.3390/ijms23073702

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

© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

The effect of IL-33 on MCPyV promoter activity in MCC-13 cells. (A) Cells were transiently co-transfected with either IL-33 expression plasmid or empty expression vector pcDNA3.1, and luciferase reporter plasmid containing either the MCPyV early or the late promoter. Luciferase activity was measured 24 h after transfection. (B) MCC-13 cells were seeded and serum starved for 24 h. Cells were then transfected with luciferase reporter plasmid containing either the MCPyV early or the late promoter, and were exposed to FL-rhIL-33 (1.25–5.0 ng/mL), CyD-rhIL-33 (1.25–5.0 ng/mL), or vehicle (PBS) for 4 h before they were harvested and luciferase activity was measured. (C) Schematic presentation of the plasmids expressing the full-length or truncated forms of the IL-33 protein. Human IL-33 mRNA (2.7 kb) encodes a protein of 270 residues with three functional domains: a nuclear, a central activation domain, and an IL-1-like cytokine domain. Sequences of the different IL-33 domains were cloned into pCMV_His(N)_FLAG(C) plasmid. (D) MCC-13 cells were transiently co-transfected with either MCPyV early and late promoter and 250 ng or 500 ng of plasmids expressing different domains of IL-33, IL-33_1–270aa, IL-33_1–65aa, IL-33_1–65,_112–270aa, IL-33_1–111aa, IL-33_66–111aa, IL-33_66–270aa, and IL-33_112–270aa. Abbreviations: FL-IL33 (full length IL-33), CyD-IL-33 (IL-33 with only cytokine domain). Each bar represents the average of three independent parallels + SD. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001. (E) MCPyV−positive WaGa cells were treated with recombinant full-length IL-33 (rhFL-IL-33) or with the cytokine domains, and expression levels of LT were compared with untreated cells. Levels of GAPDH were used as loading control. Cells were exposed for 16 h with the indicated concentrations of rhFL-IL-33 or rhCyD-Il-33. The experiment was performed two times and a representative result is shown. The ratio for empty vector:GAPDH from the two experiments was arbitrarily set as 1.0 ± standard deviation (* p < 0.05; ** p < 0.01).

The effect of IL-33 on MCPyV promoter activity in MCC-13 cells. (A) Cells were transiently co-transfected with either IL-33 expression plasmid or empty expression vector pcDNA3.1, and luciferase reporter plasmid containing either the MCPyV early or the late promoter. Luciferase activity was measured 24 h after transfection. (B) MCC-13 cells were seeded and serum starved for 24 h. Cells were then transfected with luciferase reporter plasmid containing either the MCPyV early or the late promoter, and were exposed to FL-rhIL-33 (1.25–5.0 ng/mL), CyD-rhIL-33 (1.25–5.0 ng/mL), or vehicle (PBS) for 4 h before they were harvested and luciferase activity was measured. (C) Schematic presentation of the plasmids expressing the full-length or truncated forms of the IL-33 protein. Human IL-33 mRNA (2.7 kb) encodes a protein of 270 residues with three functional domains: a nuclear, a central activation domain, and an IL-1-like cytokine domain. Sequences of the different IL-33 domains were cloned into pCMV_His(N)_FLAG(C) plasmid. (D) MCC-13 cells were transiently co-transfected with either MCPyV early and late promoter and 250 ng or 500 ng of plasmids expressing different domains of IL-33, IL-33_1–270aa, IL-33_1–65aa, IL-33_1–65,_112–270aa, IL-33_1–111aa, IL-33_66–111aa, IL-33_66–270aa, and IL-33_112–270aa. Abbreviations: FL-IL33 (full length IL-33), CyD-IL-33 (IL-33 with only cytokine domain). Each bar represents the average of three independent parallels + SD. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001. (E) MCPyV−positive WaGa cells were treated with recombinant full-length IL-33 (rhFL-IL-33) or with the cytokine domains, and expression levels of LT were compared with untreated cells. Levels of GAPDH were used as loading control. Cells were exposed for 16 h with the indicated concentrations of rhFL-IL-33 or rhCyD-Il-33. The experiment was performed two times and a representative result is shown. The ratio for empty vector:GAPDH from the two experiments was arbitrarily set as 1.0 ± standard deviation (* p < 0.05; ** p < 0.01).