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

. 2023 Nov 15;12(2):2275606. doi: 10.1080/22221751.2023.2275606

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

© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

PMC Copyright notice

Figure 2. — PLK3 interacted with NP and promoted NP phosphorylation. (A, B, C, and D) Interactions between PLK3 and viral RNP components. The expression plasmid of PLK3-Flag was respectively cotransfected with PB1-HA (A), PB2-HA (B), PA-HA (C), or NP-HA (D) in HEK293T cells. The cell lysates were collected at 24 h post-transfection. Co-IP assays were performed using an anti-Flag antibody to precipitate viral RNP components which interacted with PLK3, followed by western blotting to detect PLK3 using an anti-Flag antibody and PB1, PB2, PA, and NP using an anti-HA antibody. GAPDH served as a loading control in all of these experiments. (E) Interactions between endogenous PLK3 and viral RNP components in the context of viral infections. NPTr cells were infected with SIV-H1N1/2009 at a MOI of 0.01. The cell lysates were collected at 24 h post-infection. Co-IP assays were performed using anti-IgG and anti-PLK3 antibodies to precipitate viral RNP components which interacted with PLK3, followed by western blotting to detect PLK3 using an anti-PLK3 antibody and PB1, PB2, PA, and NP using endogenous antibodies. GAPDH served as a loading control in all of these experiments. (F, G, H, and I) Effects of PLK3 on the phosphorylation of viral RNP components. The expression plasmid of PLK3-Flag was respectively cotransfected with PB1-HA (F), PB2-HA (G), PA-HA (H), or NP-HA (I) in HEK293T cells. The cell lysates were collected at 24 h posttransfection, then, PB1-HA, PB2-HA, PA-HA, and NP-HA were immunoprecipitated with an anti-HA antibody, and the associated phosphorylated serine/threonine of PB1, PB2, PA, and NP were detected using anti-p-serine/threonine antibodies. GAPDH served as a loading control in all of these experiments.