PXD062677

PXD062677 is an original dataset announced via ProteomeXchange.

Title	Nat10 interacting proteins identified by IP and mass spectrometry (IP-MS) in mESCs
Description	The cells were cotransfected with epitope-tagged expression plasmids or empty vector(EV) . Forty-eight hours after transfection, the cells were harvested with 500 μl of lysis buffer (20 mM Tris-HCl, pH=8.0; 100 mM NaCl; 0.5% NP-40; and 1 mM EDTA) supplemented with protease inhibitor cocktail (#04693132001, Roche) and PMSF (#PB0425). The cell lysate was rotated at 4°C for 30 min. After centrifugation at 13 000 × g for 10 min, the supernatants were used for immunoprecipitation (IP). The cell extracts were incubated with anti-Flag antibody (F1804 Sigma) overnight at 4°C, followed by incubation with lysis buffer and washing with 50 μL of protein G agarose resin (Yeasen, 36405ES08) for 3 h. The incubated protein G was washed with lysis buffer at 4°C for 10 min. The cell extracts (50 μL of input) and IP samples were resuspended in 80 μL of SDS buffer, followed by pipetting up and down several times to mix the samples.For mass spectrometry analysis of heterochromatin-associated proteins and murine Nat10-associated proteins, the immunoprecipitate was subjected to 10% Acr-Bis SDS‒PAGE and separated approximately 2 cm from the well. The gel was cut and used for mass spectrometry analysis by the PTM BIO Company (Hangzhou, China).
HostingRepository	PRIDE
AnnounceDate	2025-06-23
AnnouncementXML	Submission_2025-06-22_16:05:02.533.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Guofeng Feng
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2025-04-07 03:51:56	ID requested
⏵ 1	2025-06-22 16:05:03	announced

Feng G, Yin G, Liu Y, Liu C, Li J, Lu J, Yu Y, Jin Z, Wu Y, Yue Y, Gao X, Wang J, Lu F, Liu L, Nat10 maintains the homeostasis of pluripotent and 2-cell-like states in mouse embryonic stem cells through mRNA cytidine acetylation. Nucleic Acids Res, 53(11):(2025) [pubmed]

10.1093/nar/gkaf504;

submitter keyword: IP-MS,Nat10,mESCs

Guofeng Feng
contact affiliation	Nankai University
contact email	fengguofeng_121@163.com
lab head
Guofeng Feng
contact affiliation	Nankai university
contact email	fengguofeng_121@163.com
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/06/PXD062677

PRIDE project URI

• PRIDE
  1. PXD062677
     1. Label: PRIDE project
     2. Name: Nat10 interacting proteins identified by IP and mass spectrometry (IP-MS) in mESCs