Journal: International Journal of Oral Science

Article Title: PLAGL1-IGF2 axis regulates osteogenesis of postnatal condyle development

doi: 10.1038/s41368-025-00386-4

Figure Lengend Snippet: Plagl1 deficiency leads to downregulated IGF2 and increased glucose metabolism. a Protein-protein interaction analysis for the relationship between PLAGL1 and genes activated by osteogenic induction while inhibited by knockdown of Plagl1 . b Western blot analysis of the expression of IGF2 during osteogenic induction. c Western blot analysis of the expression of IGF2 after knockdown of Plagl1 . d , e Immunofluorescence staining of IGF2 (green) and DAPI (cyan) in condyles of control and Wnt1-Cre;Plagl1 pat fl/+ mice at PN 28.5. The boxes ( d1, e1 ) show the magnified regions on the right. White arrows show the positive signals. n = 3 per group. Scale bars, 50 μm. f Western blot analysis of the expression of IGF2 in the subchondral bone after knockdown of Plagl1 . Band intensities of IGF2 normalized to β-actin. n = 3 per group. Unpaired two-tailed t-test was used for statistical analysis. GSEA plots of GO term 0006119 ( g ) and 0061621 ( h ) between AdV-EGFP and AdV-CRE-EGFP transfected condyle mesenchymal cells after 4 days of osteogenic induction. n = 3 per group. Analysis of glucose uptake ( i ), intracellular ATP levels ( j ), oxygen consumption rate ( k ), reactive oxygen species ( l ), and L-lactate production ( m ) in AdV-EGFP and AdV-CRE-EGFP transfected condyle mesenchymal cells after 4 days of osteogenic induction. n = 4 per group for i. n = 3 per group for j, k, m. n = 9 per group for l. Unpaired two-tailed t-test was used for statistical analysis. The results are presented as the mean ± standard deviation (SD). * P < 0.05. ns, no significant difference

Article Snippet: To confirm that IGF2 is a downstream target of PLAGL1 during osteoblastic differentiation, we added recombinant human IGF2 (Novoprotein, CF61) at 250 ng/mL in Plagl1 -deficient cells during osteogenic induction.

Techniques: Knockdown, Western Blot, Expressing, Immunofluorescence, Staining, Control, Two Tailed Test, Transfection, Standard Deviation

Journal: International Journal of Oral Science

Article Title: PLAGL1-IGF2 axis regulates osteogenesis of postnatal condyle development

doi: 10.1038/s41368-025-00386-4

Figure Lengend Snippet: IGF2 administration partially rescues the osteoblastic differentiation of Plagl1 -deficient cells. qPCR for the expression of osteoblast-linked gene Col1α1 ( a ) and Dmp1 ( b ) with the addition of IGF2 in AdV-EGFP and AdV-CRE-EGFP transfected condyle mesenchymal cells. n = 3 per group. One-way analysis of variance with Tukey’s post-hoc test was used for statistical analysis. c Western blot analysis of the expression of osteoblast-linked protein RUNX2, COl1α1, and DMP1 with the addition of IGF2 in AdV-EGFP and AdV-CRE-EGFP transfected condyle mesenchymal cells. d Immunofluorescence staining of rescued RUNX2 (red) and DAPI (cyan) in AdV-EGFP and AdV-CRE-EGFP transfected condyle mesenchymal cells with the addition of IGF2 or control vector. White arrows showed the positive signals. n = 3 per group. Scale bars, 250 μm. e ALP staining showing osteoblastic capacity in AdV-EGFP and AdV-CRE-EGFP transfected condyle mesenchymal cells with the addition of IGF2 or control vector. n = 3 per group. Scale bars, 200 μm. The results are presented as the mean ± standard deviation (SD). * P < 0.05. ns, no significant difference

Article Snippet: To confirm that IGF2 is a downstream target of PLAGL1 during osteoblastic differentiation, we added recombinant human IGF2 (Novoprotein, CF61) at 250 ng/mL in Plagl1 -deficient cells during osteogenic induction.

Techniques: Expressing, Transfection, Western Blot, Immunofluorescence, Staining, Control, Plasmid Preparation, Standard Deviation

Journal: International Journal of Oral Science

Article Title: PLAGL1-IGF2 axis regulates osteogenesis of postnatal condyle development

doi: 10.1038/s41368-025-00386-4

Figure Lengend Snippet: Schematic diagram of the PLAGL1-IGF2 axis in mandibular condylar development. Under physiological conditions, PLAGL1 activates Igf2 expression to sustain the IGF2/IGFBP signaling pathway, thereby maintaining glucose metabolic homeostasis and promoting osteoblast differentiation through upregulation of key osteogenic markers of Sp7 , Runx2 , Col1α1 and Dmp1 . In contrast, Plagl1 deficiency suppresses IGF2 signaling, triggering metabolic dysregulation characterized by aberrantly elevated oxidative phosphorylation and glycolysis, which collectively impair osteogenic differentiation and ultimately disrupt mandibular condylar morphogenesis

Article Snippet: To confirm that IGF2 is a downstream target of PLAGL1 during osteoblastic differentiation, we added recombinant human IGF2 (Novoprotein, CF61) at 250 ng/mL in Plagl1 -deficient cells during osteogenic induction.

Techniques: Expressing, Phospho-proteomics