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The lactylome of mice granulosa cells

ABSTRACT: Follicular GCs were obtained through intraperitoneal injections of 10, 5, 2, and 2 IU FSH at 12 h intervals. For the enrichment of Kla-modified peptides, the tryptic peptides were solubilized in NETN buffer (1 mM EDTA, 50 mM Tris–HCl, 100 mM NaCl, 0.5% NP-40, pH 8.0) and then incubated with prewashed beads (PTM Bio) at 4 °C for 12 h. Elution of the bound peptides was achieved using trifluoroacetic acid (0.1%). The eluted peptides were subsequently vacuum-dried and subjected to desalting using C18 ZipTips (Millipore) prior to LC‒MS/MS analysis.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Ovary

SUBMITTER: Gang Wu

LAB HEAD: GANG WU

PROVIDER: PXD059168 | Pride | 2025-09-29

REPOSITORIES: Pride

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Publications

Lactylation of CREB is required for FSH-induced proliferation and differentiation of ovarian granulosa cells.

Wu Gang G Chen Min M He Tong T Pan Yitong Y Li Chengyu C Liu Zhaojun Z Li Hongmin H Sheng Yanan Y Dai Weilong W Shen Ming M Liu Honglin H

Nucleic acids research 20250901 17

Follicle-stimulating hormone (FSH) promotes follicular development by inducing the proliferation and differentiation of granulosa cells (GCs). This process is primarily attributed to the activation of the canonical G protein-coupled receptor (GPCR)/adenylyl cyclase/cAMP/PKA/CREB signaling pathway. Here, we revealed a novel mechanism wherein FSH promotes GCs proliferation and differentiation by stimulating cAMP response element-binding protein (CREB) lactylation. Specifically, FSH induced CREB la ...[more]

PMID: 40966521

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Lactylome and proteomics analyses of bladder cancer

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Quantification of lactylation in mouse peritoneal macrophages with or without stimulation of VSV by LS-MS/MS

Project description:Quantification of lactylation in mouse peritoneal macrophages with or without stimulation of VSV(12h) by LS-MS/MS, And lactylation peptides were enriched by Anti-Lactyllysine antibody conjugated agarose beads (PTM-1404, PTM Bio)

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Gene expression in bovine ovarian follicle granulosa

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PATCH labeling in the BSA through red light

Project description:For the liquid chromatography-mass spectrometry (LC-MS) analysis, the 500 μM AP was used to label BSA in the reaction, followed by clicking chemical conjugation with 0.1 mM azide-PEG3-biotin under 0.5 mM CuCl(2), 2 Mm Tris (3-hydroxypropyltriazolylmethyl) amine (THPTA, Sigma-Aldrich) and 5 mM ascorbic acid. Then, the identification of the labeled site was analyzed by a HF-X mass spectrometer (ThermoFisher) that was coupled to a nano Elute liquid chromatography system (Bruker Daltonics). MS data were processed by the Byonic software 4.0.12 version (Protein Metrics INC.) for both de novo sequencing and database search. The peptides with scores higher than 200 were retained for analysis.

2025-07-30 | PXD057016 | Pride

Qualitative LC-MSMS analysis of Mus Musculus proteins

Project description:30 µg proteins per sample were reduced (20 mM DTT Sigma, room temperature RT, 1 h) and S-alkylated (50 mM IAA Sigma, 1 h, dark). The remaining IAA was quenched with 20 mM DTT for 1 h in the dark. The proteins were digested with 1:50(w/w enzyme:protein) MS-grade trypsin/lys-C mix (Thermo Scientific) for 24 h at RT. The enzyme digestion was quenched by lowering the pH with formic acid (Fisher) and the sample was desalted with ZipTip C18 (Merck-Millipore) column. Samples were resuspended in 10 µl of 0.2% formic acid prior LC-MS/MS analysis.

2025-05-06 | PXD051730 | Pride

Analysis of Lysine Lactylation in CD4+ T cells of normal and EAU mouse

Project description:Primary CD4+ T cells were isolated from normal and EAU mice spleens and used for lactylome analysis.

2023-08-29 | PXD040514 | Pride

H3K27me3 in Phytophthora sojae P6497 and its pssu(z)12 mutant at mycelium stage

Project description:We report the H3K27me3 profile on Avr1b locus in two Phytophthora sojae strains P6497 and pssu(z)12 mutant. Nuclei of Phytophthora sojae P6497 and pssu(z)12 mutant T34 (lost 561bp by CRISPR/Cas9) mycelium (3-days old) was extracted and digested to 200-400bp using micrococcal nuclease (MNase: NEB M0247S). The antibody Millipore 07-449 was used to immunoprecipitation. We find significant accumulation of H3K27me3 at Avr1b locus in Avr1b silencing strain P6497 and clear H3K27me3 depletion at Avr1b locus in Avr1b unsilenced strain pssu(z)12 mutant T34.

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Identification of type strain-specific diagnostic peptides of the Bacillus cereus group

Project description:The Bacillus cereus group consists of eight very closely related species and comprises both harmless and human pathogenic species. Yet, methods to rapidly and accurately distinguish these species are currently lacking as we demonstrate that classical matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) biotyping did not achieve reliable identification of each type strain. We assigned type strain-specific diagnostic peptides to the B. cereus group based on comparisons of their proteome profiles. The number of diagnostic peptides varies remarkably in type strain-dependent manner. The state of the art of the reference database is crucial in the process of validating candidate diagnostic peptides and may lead to a noteworthy reduction of verified diagnostic peptides as putative diagnostic peptides might be found in other species as well.

2018-02-07 | PXD003669 | Pride

Differential protein expression between Carfilzomib-sensitive (AMO1) and resistant (AMO1-CFZ) multiple myeloma cells

Project description:Carfilzomib-sensitive (AMO1) and resistant (AMO1-CFZ) MM cells were grown in RPMI-1640 medium supplemented with 10 % FBS and 0.68 mM L-glutamine, in a humidified incubator at 5% CO2 and 37 °C. The AMO1-CFZ medium was additionally added 90 nM CFZ, while the CFZ sensitive AMO1 cells were added vehicle (0.009 % DMSO) only. AMO1-CFZ was either harvested directly or grown for 1 week in CFZ free medium (vehicle only) prior to harvest. AMO1 and carfilzomib-resistant AMO1-CFZ cells were resuspended in 100 µl 1% sodium deoxycholate, 100 mM Tris-HCl pH 8.5, 10 mM tris(2-carboxyethyl)phosphine (TCEP), 40 mM chloroacetamide (CAA), heated at 90 °C for 45 min and sonicated for 10 cycles (30 s ON/30 s OFF) using a Bioruptor pico sonicator. After centrifugation at 16000 × g for 10 min, 50 µg soluble protein from each sample was added 100 µl 0.1M ammonium bicarbonate, 0.5 µg trypsin and digested overnight at 37°C. Peptides were desalted using C18 spin columns, dried in a speedvac centrifuge and resuspended in 0.1% formic acid prior to MS analysis. Label-free quantitatative (LFQ) LC-MS/MS was performed on a timsTOF Pro (Bruker Daltonics) connected to a nanoElute (Bruker Daltonics) HPLC. Peptides were separated over a Bruker PepSep C18 (75 µm × 15 cm) column with running buffers A (0.1 % formic acid) and B (0.1 % formic acid in acetonitrile) using a 100 min gradient from 2 % B to 40 % B. The timsTof instrument was operated in the DDA PASEF mode with 10 PASEF scans per acquisition cycle and accumulation and ramp times of 100 ms each. The ‘target value’ was set to 20000 and dynamic exclusion was activated and set to 0.4 min. The quadrupole isolation width was set to 2 Th for m/z < 700 and 3 Th for m/z > 800.

2024-06-07 | PXD045439 | Pride

Gene expression in bovine ovarian follicle granulosa

Project description:Granulosa cells mature and die as ovarian follicles enlarge and die (undergo atresia) under the influence of hormones and intrafollicular factors. Later in follicular development, a fluid-filled antrum is formed, a process which is accompanied by a high rate of atresia. These small antral follicles (5 mm or less in diameter in the cow) contain granulosa of 2 different phenotypes, rounded or columnar, whereas follicles larger than 5 mm have the rounded phenotype only. Prior to ovulation, in larger follicles greater than 10 mm in size, the granulosa begin to migrate and differentiate in preparation for oocyte release and formation of the corpus luteum. These two key phases of follicular development were studied by gene expression microarray analysis using a bovine model to dissect the molecular mechanisms underlying these processes. Four groups of bovine ovarian follicles were selected for analysis. Follicle size, type and array number for each group are as follows: small (3-5 mm) healthy rounded (n=5), small healthy columnar (n=5), atretic (n=5) and large healthy (>10 mm; n=4). For each group, the RNA from a single follicle was used to hybridise an array, except for 3 small healthy samples which were pooled from 2 follicles each due to low RNA.

2014-05-20 | E-GEOD-39589 | biostudies-arrayexpress