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 19;12:2954. doi: 10.1038/s41467-021-23264-z

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. 1 — a Second structure of MIF substrates ssDNA with the stem-loop (SL) and Y-shaped dsDNA with different lengths of unpaired nucleotides at the 3′ end. b, c In vitro MIF (2 μM) nuclease cleavage assay using Y-shaped dsDNA as substrates. Data were quantified in c (mean ± SEM, n = 3 biologically independent experiments). ****P < 0.0001, ***P < 0.001; by one-way ANOVA Dunnett’s multiple comparisons test vs 0 nt group. EtBr ethidium bromide. d, e In vitro MIF (2 μM) nuclease cleavage assay using 3′ biotin-labeled Y-shaped dsDNA as substrates. Data were quantified in e (mean ± SEM, n = 3 biologically independent experiments). ****P < 0.0001, *P < 0.05; by one-way ANOVA Dunnett’s multiple comparisons test vs 7b group. Red dot indicates biotin (d). Arrow indicates the cleavage products. f Protein purification of wild-type (WT) MIF and nuclease-inactive E22A mutant shown by Coomassie blue staining. BSA (0.5–3 μg) was used as control. g In vitro nuclease assay of MIF and E22A mutant (2 μM) using 3′ biotin-labeled Y-shaped dsDNA (0.8 μM) as substrates. h, i MIF (2 μM) cleaves DS-1 nt substrate (0.8 μM) in a time (1, 5, 10, 15, 30, 60 min)-dependent manner. Representative image is shown in h. Data are quantified in i (mean ± SEM, n = 3 biologically independent experiments). j, k MIF (2 μM) cleaves DS-1 nt substrate in a concentration (0.2, 0.4, 1, 2, 4 μM)-dependent manner. Representative image is shown in j. Data are quantified in k (mean ± SEM, n = 3 biologically independent experiments).