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 Jan 28;71(6):1815–1827. doi: 10.1093/jxb/erz542

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

© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Fig. 5. — Mutation of the woolly motif attenuates the interaction between Nbwo and NbCycB2 in N. benthamiana. (a) Yeast two-hybrid (Y2H) assays of the interactions between NbCycB2 and Nbwo or NbWo^V using QDO/X/A medium. Clones with pGADT7-53 (BD-53) and pGADT7-T (AD-T) served as positive controls, and clones with pGBKT7-Lam (BD-Lam) and pGADT7-T (AD-T) served as negative controls. Only the positive control and AD-Nbwo and BD-NbcycB2 could grow on the medium. (b) Pull-down assays between Nbwo or NbWo^V with NbCycB2 proteins. The NbCycB2-GST protein was immunoprecipitated with glutathione agarose, and the immunoblots were probed with anti-HIS and anti-GST antibodies. Only the recombinant HIS-Nbwo protein could co-precipitate with GST-NbCycB2. The asterisks indicate the bands for Nbwo or NbWo^V. (c) Y3H assays to determine the competition between NbCycB2 and the LZ domain of Nbwo for binding to Nbwo (fused to GAL4 DNA-AD). The methionine-repressible promoter in the pBridge vector controlled the expression of NbCycB2 in the presence of the Nbwo LZ domain (fused to GAL4 DNA-BD). (d) Pull-down assays to determine whether NbCycB2 could compete for binding to Nbwo. The total protein from P35S::HA-Nbwo protoplasts was immunoprecipitated with anti-HA beads, and the immunoblots were probed with anti-Flag and anti-MBP antibodies. (This figure is available in colour at JXB online.)