Dataset Information

Sam68 interactome in mouse embryonic stem cells (mESCs)

ABSTRACT: Pellets from a mESC line with GFP tagged Sam68 and a control line (mESC expressing GFP alone) were collected at different time points during differentiation (Day 0, Day 3, Day 10).Pull-down was performed from normalized total cell lysates with GFP-trap beads (Chromotek) and analyzed by mass spectrometry.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Embryo, Stem Cell

SUBMITTER: Giulia Calloni

LAB HEAD: R. Martin Vabulas

PROVIDER: PXD009209 | Pride | 2026-06-08

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	180208_gc_MCA-Sam68_PD_10A.raw	Raw
	180208_gc_MCA-Sam68_PD_10B.raw	Raw
	180208_gc_MCA-Sam68_PD_10C.raw	Raw
	180208_gc_MCA-Sam68_PD_11A.raw	Raw
	180208_gc_MCA-Sam68_PD_11B.raw	Raw

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Publications

SAM68 is a multifunctional post-transcriptional regulator of cardiomyocyte differentiation.

Broglia Laura L Dasti Alessandro A Antonelli Maria Carla MC Aoun Guy G D'Agostino Sabrina S Vandelli Andrea A Armaos Alexandros A Delli Ponti Riccardo R Wolf Sarah S Klostermann Melina M Arnal Segura Magdalena M Tian Tian V TV Mariani Davide D Colantoni Alessio A Paronetto Maria Paola MP Gustincich Stefano S Zarnack Kathi K Bechara Elias E Tartaglia Gian Gaetano GG

Nucleic acids research 20260501 10

RNA-binding proteins (RBPs) of the STAR family play key roles in mammalian development, yet their contributions to lineage specification remain incompletely understood. Here, using CRISPR-Cas9 knockout models combined with multi-omics approaches, we investigate the functions of two STAR proteins, SAM68 and QKI, in mouse embryonic stem cells (mESCs). Both RBPs support mESC proliferation, self-renewal, and efficient differentiation into cardiomyocytes. Although SAM68 and QKI belong to the same pro ...[more]

PMID: 42240622

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Project description:Cardiac disease accounts for the largest proportion of adult mortality and morbidity in the industrialized world. However, progress toward improved clinical treatments is hampered by an incomplete understanding of the genetic programs controlling early cardiogenesis. To better understand this process, we set out to identify genes whose expression is enriched within early cardiac fated populations, obtaining the transcriptional signatures of mouse embryonic stem cells (mESCs) differentiating along a cardiac path. We compared the RNA profiles of cardiac precursors cells (CPCs) with time-matched non-CPCs and undifferentiated mESCs, using a transgenic mESC line harboring an Nkx2-5 cardiac-specific regulatory sequence driving green fluorescent protein (GFP) to facilitate selection of CPCs. Approximately 24% (43/176) of the transcripts enriched in the CPC population have known roles in cardiac function or development. Importantly, we evaluated the biological relevance of a subset 31/133 (23%) of the remaining candidate genes by in situ hybridization and report that all were expressed in key cardiac structures during cardiogenesis (embryonic day, E7.5 - 9.5), many of which were previously uncharacterized. These data demonstrate the power of mESC differentiation to model specific developmental processes and provide a valuable resource that may be mined to further elucidate the genetic programs underlying cardiogenesis. Experiment Overall Design: Transgenic mouse embryonic stem cell line (Nkx2-5-GFP, PMID:9834187) contains an Nkx2-5 cardiac-specific transcription factor driving expression of green fluorescent protein (GFP). Cells were differentiated through embryoid body formation (hanging droplet technique) as described in PMID:8155574. Following primary mouse embryonic fibroblasts (PMEF) feeder subtraction, mESCs were dissociated and resuspended in differentiation medium. Cells at this time point were designated as day 0. Following incubation for two days, the EBs were transferred, in suspension, to poly-HEMA coated tissue culture dishes to prevent cell attachment and grown in medium containing ascorbic acid PMID:12668514) which was replenished every 3 days thereafter. Experiment Overall Design: Cells were harvested on days 0, 4 and 6 in three biological replicates Experiment Overall Design: each (1a, 2a and 3a), washed in PBS for 30 minutes at 37ÂºC then resuspended in 0.25% trypsin for 7 minutes. Experiment Overall Design: Cells from day 6 were then sorted using the BD Biosciences FACSAriaâ¢ Cell sorting system into GFP+ and GFP- fractions into RLT buffer (RNeasy, Qiagen). This sorting separates cells into two groups reflecting their differentiation into those fated to a non-cardiac lineage (GFP negative) and to a cardiac lineage (GFP positive).