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

. 2021 May 12;12:2743. doi: 10.1038/s41467-021-22733-9

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

© The Author(s) 2021

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. 4 — a Interaction of GST-CTD with His₆-SUMO-INI1_183-304. A titration curve was generated with increasing concentrations of His₆-SUMO-INI1_183-304 with two different fixed concentration of GST-CTD and the interactions were detected as Alpha Score. The K_D values were determined by nonlinear regression analysis using specific binding with Hill slope analysis in GraphPad Prism. Data from one representative experiment is depicted. b, c Effect of salt on the interactions of GST-CTD either with His₆-SUMO-INI1_183-304 or biotinylated(Bio)-TAR RNA (n = 3 independent experiments). The interaction was tested using fixed concentrations of GST-CTD (0.75 μM) and increasing concentrations of His₆-SUMO-INI1_183-304 (or Bio-TAR RNA) in two different NaCl conditions (100 and 500 mM); d, e Inhibition of GST-CTD interaction with His₆-SUMO-INI1_183-304 or Bio-TAR RNA (n = 6 independent experiments). Interactions were set up between GST-CTD (0.186 μM) with His₆-SUMO-INI1_183-304 (0.094 μM), or GST-CTD (0.03 μM) with Bio-TAR RNA (0.1 μM) and increasing concentration of the third component (indicated in the X-axis) was added to the reaction. The IC₅₀ values were determined by fitting the data to a four-parameter dose-response curve using GraphPad prism. f–h Interaction of GST-CTD and its substitution mutants with INI1_183-304 and TAR RNA (n = 3 independent experiments). Representative Coomassie gel (from one out of three independent experiments) showing equal loading of the wild type and mutant proteins for the binding assays (f) and uncropped gels are provided in the source data. Interaction of GST-CTD and mutants with INI1_183-304 (g), and with biotinylated-TAR RNA (h). The graphs represent the % of the interaction of mutants as compared to that of wild type (WT) IN set at 100%. For both panels (g) and (h), WT and mutants are represented in different colors as indicated in the key provided next to the bar graphs. In all panels, except in a, graphs represent Mean ± SEM. In all panels the pink cartoon Rpt1 represents INI1_183-304, blue cartoon CTD, IN-CTD, and red stem-loop, TAR RNA.