Related Experiment Video

Updated: Feb 22, 2026

Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis
07:26

Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis

Published on: April 1, 2022

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Integrated Phosphoproteomics Identifies TGFβ-Dependent Phosphorylation Events Linking Kinase Signaling to Autophagy

Xia Peng1, Jing Chen1,2, Xiaoyu Zheng1

  • 1Laboratory of Orofacial Development, Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Beijing Stomatological Hospital, Capital Medical University, No. 9 Fanjiacun Road, FengTai District, Beijing 100070, China.

Proteomes
|February 20, 2026
PubMed
Abstract

Insights

Transforming the understanding of cleft palate development, this study reveals how TGFβ2 signaling influences cell phosphorylation. Autophagy pathways are key players in regulating palatal mesenchyme cells during embryonic development.

Area of Science:

  • Craniofacial development research
  • Cell signaling pathways
  • Molecular biology

Background:

  • Cleft palate (CP) is a common craniofacial birth defect.
  • The transforming growth factor beta (TGFβ) pathway is crucial in palate development.
  • The role of autophagy in regulating mouse embryonic palatal mesenchyme (MEPM) cells and its interaction with TGFβ signaling is not well understood.

Purpose of the Study:

  • To investigate the interplay between autophagy and TGFβ signaling during palatogenesis.
  • To elucidate the molecular mechanisms underlying TGFβ-induced phosphoproteome changes in MEPM cells.

Main Methods:

  • Histological analyses (H&E, IHC) to assess protein expression during palatal development.
  • Bulk RNA sequencing (Bulk RNA-seq) to identify enriched pathways.
  • TMT-based phosphoproteomics to analyze protein phosphorylation in TGFβ2-treated MEPM cells.

Main Results:

  • Increased expression of autophagy markers (Beclin 1, LC3) during critical palatal fusion stages.
  • Bulk RNA-seq revealed enrichment of autophagy and TGFβ signaling pathways.
  • Phosphoproteomics identified numerous differentially abundant phosphopeptides and enriched phosphorylation motifs, highlighting CSNK2A, MAPKs, and CDKs as key regulators.
  • Functional enrichment analysis implicated mTOR, MAPK, and autophagy/mitophagy pathways.

Conclusions:

  • TGFβ2 significantly alters the MEPM cell phosphoproteome via a Smad-independent pathway.
  • This study expands the knowledge of palate-specific phosphosignaling beyond the canonical Smad cascade.
  • Identified novel regulators and pathways involved in TGFβ-mediated palatogenesis.
Keywords:
TGFβ2autophagymouse embryonic palatal mesenchyme (MEPM) cellpalatogenesisphosphoproteomics

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