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 Apr 19;49(9):5003–5016. doi: 10.1093/nar/gkab269

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

© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

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

Figure 1. — Depletion of LIG1 inhibits Okazaki fragment ligation in Xenopus egg extracts. (A) Control or xLIG1 antibodies were used for immunoprecipitation (I. P.) and immunoprecipitated proteins were analyzed by immunoblotting using the indicated antibodies (n = 2). (B) Sequences of the wild-type and the F8A/F9A mutant PIP-box in xLIG1 are shown. (C) Recombinant wild-type xLIG1–3 × Flag or xLIG1-F8A/F9A-3 × Flag was added to the egg extracts. Immunoprecipitation using anti-Flag antibodies was performed and resultant immunoprecipitates were analyzed by immunoblotting using the indicated antibodies (n = 2). (D) Interphase egg extracts were added with sperm chromatin and incubated in the presence of 30 μM aphidicolin or in its absence (DMSO). Chromatin fractions were isolated and analyzed by immunoblotting using the indicated antibodies (n = 2). (E) Immunodepletion efficiency of xLIG1 from Xenopus egg extract (n = 3). (F) Replication products in mock- or xLIG1-depleted extracts were separated by alkaline agarose gel electrophoresis followed by autoradiography (n = 2). (G) xLIG1-depleted extracts were supplemented with wild-type xLIG1–3 × Flag or xLIG1-K721A-3 × Flag and chromatin was isolated. Purified genomic DNA from chromatin was labeled using exonuclease-deficient Klenow fragment and α-³²P dCTP, and separated in a denaturing agarose gel (n = 2).