Dataset Information

Nat10 interacting proteins identified by IP and mass spectrometry (IP-MS) in mESCs

ABSTRACT: 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).

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Embryonic Stem Cell

SUBMITTER: Guofeng Feng

LAB HEAD: Guofeng Feng

PROVIDER: PXD062677 | Pride | 2025-06-23

REPOSITORIES: Pride

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Publications

Nat10 maintains the homeostasis of pluripotent and 2-cell-like states in mouse embryonic stem cells through mRNA cytidine acetylation.

Feng Guofeng G Yin Guoxing G Liu Yusheng Y Liu Chang C Li Jie J Lu Jiangtao J Yu Yongqin Y Jin Ziyi Z Wu Yiwei Y Yue Yanbin Y Gao Xiufang X Wang Jiaqiang J Lu Falong F Liu Lin L

Nucleic acids research 20250601 11

Naïve mouse embryonic stem cells (mESCs) are characterized by a mixed population of cells in an interconvertible pluripotent state and a totipotent 2-cell (2C)-like state. It remains to be understood how the pluripotent state is maintained while the 2C-like state is suppressed. We show that N-acetyltransferase 10 (Nat10) maintains the pluripotent state and suppresses the 2C-like state in mESCs through mRNA modification and stabilization. Nat10 as a nucleolar protein may indirectly interact with ...[more]

PMID: 40521666

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			Action	DRS
	FLAG_EV.raw	Raw
	FLAG_EV.xlsx	Xlsx
	FLAG_NAT10.raw	Raw
	FLAG_NAT10.xlsx	Xlsx