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( A ) scRNA-seq workflow. Created in BioRender. Kadota, C. (2026) https://BioRender.com/ks47d2b . ( B ) UMAP visualization of cell types in TMJ. Chondrocytes are highlighted in blue. ( C ) Proportion of each cell cluster in p75 +/+ and p75 −/− MCCs. ( D ) Pathway analysis of chondrocytes identified by scRNA-seq. ( E ) <t>Stereomicroscope</t> images and gross pellet size. Scale bar, 500 μm. ( F ) Safranin O staining [Top: Whole-pellet view (10×; scale bar, 500 μm); bottom, 40× view of the dotted area. Scale bar, 100 μm] and quantification of Safranin O–positive area, and ( G ) gene expression analyses ( Acan , Ihh , Sox9 , and Col2a1 ) of p75 +/+ and p75 −/− mBMSCs underwent 3D pellet culture ( N = 6 per group). ( H ) Left: Western blotting of p75 NTR , phosphorylated-AKT, AKT, phosphorylated-p38, and p38 in mBMSCs treated with vehicle or BDNF. Right: Quantification of p-AKT/AKT and p-p38/p38 ratios. ( N = 5 independent experiments). ( I ) Gene expression analyses in p75 +/+ and p75 −/− condyles for p75 NTR , Ntrk2 , Bdnf , Trp53 , Bax , Acan , Ihh , Sox9 , Col2a1 , and Mki67 ( N = 5 per group). IF staining for ( J ) Ki-67 and ( K ) TUNEL and quantification of Ki-67 and TUNEL-positive cells in p75 +/+ and p75 −/− TMJ ( N = 5 per group). Blue, DAPI. Scale bar, 50 μm. ( L ) Cell proliferation assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). ( M ) Apoptosis assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). Statistical significance was determined using unpaired two-sided Student’s t test or two-way ANOVA, as appropriate. Bonferroni correction was applied for pairwise post hoc comparisons following two-way ANOVA. Results are represented as the means ± 1 SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. N represents the number of mice or experimental replicates.
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( A ) scRNA-seq workflow. Created in BioRender. Kadota, C. (2026) https://BioRender.com/ks47d2b . ( B ) UMAP visualization of cell types in TMJ. Chondrocytes are highlighted in blue. ( C ) Proportion of each cell cluster in p75 +/+ and p75 −/− MCCs. ( D ) Pathway analysis of chondrocytes identified by scRNA-seq. ( E ) <t>Stereomicroscope</t> images and gross pellet size. Scale bar, 500 μm. ( F ) Safranin O staining [Top: Whole-pellet view (10×; scale bar, 500 μm); bottom, 40× view of the dotted area. Scale bar, 100 μm] and quantification of Safranin O–positive area, and ( G ) gene expression analyses ( Acan , Ihh , Sox9 , and Col2a1 ) of p75 +/+ and p75 −/− mBMSCs underwent 3D pellet culture ( N = 6 per group). ( H ) Left: Western blotting of p75 NTR , phosphorylated-AKT, AKT, phosphorylated-p38, and p38 in mBMSCs treated with vehicle or BDNF. Right: Quantification of p-AKT/AKT and p-p38/p38 ratios. ( N = 5 independent experiments). ( I ) Gene expression analyses in p75 +/+ and p75 −/− condyles for p75 NTR , Ntrk2 , Bdnf , Trp53 , Bax , Acan , Ihh , Sox9 , Col2a1 , and Mki67 ( N = 5 per group). IF staining for ( J ) Ki-67 and ( K ) TUNEL and quantification of Ki-67 and TUNEL-positive cells in p75 +/+ and p75 −/− TMJ ( N = 5 per group). Blue, DAPI. Scale bar, 50 μm. ( L ) Cell proliferation assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). ( M ) Apoptosis assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). Statistical significance was determined using unpaired two-sided Student’s t test or two-way ANOVA, as appropriate. Bonferroni correction was applied for pairwise post hoc comparisons following two-way ANOVA. Results are represented as the means ± 1 SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. N represents the number of mice or experimental replicates.
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( A ) scRNA-seq workflow. Created in BioRender. Kadota, C. (2026) https://BioRender.com/ks47d2b . ( B ) UMAP visualization of cell types in TMJ. Chondrocytes are highlighted in blue. ( C ) Proportion of each cell cluster in p75 +/+ and p75 −/− MCCs. ( D ) Pathway analysis of chondrocytes identified by scRNA-seq. ( E ) <t>Stereomicroscope</t> images and gross pellet size. Scale bar, 500 μm. ( F ) Safranin O staining [Top: Whole-pellet view (10×; scale bar, 500 μm); bottom, 40× view of the dotted area. Scale bar, 100 μm] and quantification of Safranin O–positive area, and ( G ) gene expression analyses ( Acan , Ihh , Sox9 , and Col2a1 ) of p75 +/+ and p75 −/− mBMSCs underwent 3D pellet culture ( N = 6 per group). ( H ) Left: Western blotting of p75 NTR , phosphorylated-AKT, AKT, phosphorylated-p38, and p38 in mBMSCs treated with vehicle or BDNF. Right: Quantification of p-AKT/AKT and p-p38/p38 ratios. ( N = 5 independent experiments). ( I ) Gene expression analyses in p75 +/+ and p75 −/− condyles for p75 NTR , Ntrk2 , Bdnf , Trp53 , Bax , Acan , Ihh , Sox9 , Col2a1 , and Mki67 ( N = 5 per group). IF staining for ( J ) Ki-67 and ( K ) TUNEL and quantification of Ki-67 and TUNEL-positive cells in p75 +/+ and p75 −/− TMJ ( N = 5 per group). Blue, DAPI. Scale bar, 50 μm. ( L ) Cell proliferation assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). ( M ) Apoptosis assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). Statistical significance was determined using unpaired two-sided Student’s t test or two-way ANOVA, as appropriate. Bonferroni correction was applied for pairwise post hoc comparisons following two-way ANOVA. Results are represented as the means ± 1 SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. N represents the number of mice or experimental replicates.
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( A ) scRNA-seq workflow. Created in BioRender. Kadota, C. (2026) https://BioRender.com/ks47d2b . ( B ) UMAP visualization of cell types in TMJ. Chondrocytes are highlighted in blue. ( C ) Proportion of each cell cluster in p75 +/+ and p75 −/− MCCs. ( D ) Pathway analysis of chondrocytes identified by scRNA-seq. ( E ) Stereomicroscope images and gross pellet size. Scale bar, 500 μm. ( F ) Safranin O staining [Top: Whole-pellet view (10×; scale bar, 500 μm); bottom, 40× view of the dotted area. Scale bar, 100 μm] and quantification of Safranin O–positive area, and ( G ) gene expression analyses ( Acan , Ihh , Sox9 , and Col2a1 ) of p75 +/+ and p75 −/− mBMSCs underwent 3D pellet culture ( N = 6 per group). ( H ) Left: Western blotting of p75 NTR , phosphorylated-AKT, AKT, phosphorylated-p38, and p38 in mBMSCs treated with vehicle or BDNF. Right: Quantification of p-AKT/AKT and p-p38/p38 ratios. ( N = 5 independent experiments). ( I ) Gene expression analyses in p75 +/+ and p75 −/− condyles for p75 NTR , Ntrk2 , Bdnf , Trp53 , Bax , Acan , Ihh , Sox9 , Col2a1 , and Mki67 ( N = 5 per group). IF staining for ( J ) Ki-67 and ( K ) TUNEL and quantification of Ki-67 and TUNEL-positive cells in p75 +/+ and p75 −/− TMJ ( N = 5 per group). Blue, DAPI. Scale bar, 50 μm. ( L ) Cell proliferation assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). ( M ) Apoptosis assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). Statistical significance was determined using unpaired two-sided Student’s t test or two-way ANOVA, as appropriate. Bonferroni correction was applied for pairwise post hoc comparisons following two-way ANOVA. Results are represented as the means ± 1 SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. N represents the number of mice or experimental replicates.

Journal: Science Advances

Article Title: p75 NTR orchestrates chondrogenesis and mechanoadaptation to preserve temporomandibular joint development and homeostasis

doi: 10.1126/sciadv.aef7786

Figure Lengend Snippet: ( A ) scRNA-seq workflow. Created in BioRender. Kadota, C. (2026) https://BioRender.com/ks47d2b . ( B ) UMAP visualization of cell types in TMJ. Chondrocytes are highlighted in blue. ( C ) Proportion of each cell cluster in p75 +/+ and p75 −/− MCCs. ( D ) Pathway analysis of chondrocytes identified by scRNA-seq. ( E ) Stereomicroscope images and gross pellet size. Scale bar, 500 μm. ( F ) Safranin O staining [Top: Whole-pellet view (10×; scale bar, 500 μm); bottom, 40× view of the dotted area. Scale bar, 100 μm] and quantification of Safranin O–positive area, and ( G ) gene expression analyses ( Acan , Ihh , Sox9 , and Col2a1 ) of p75 +/+ and p75 −/− mBMSCs underwent 3D pellet culture ( N = 6 per group). ( H ) Left: Western blotting of p75 NTR , phosphorylated-AKT, AKT, phosphorylated-p38, and p38 in mBMSCs treated with vehicle or BDNF. Right: Quantification of p-AKT/AKT and p-p38/p38 ratios. ( N = 5 independent experiments). ( I ) Gene expression analyses in p75 +/+ and p75 −/− condyles for p75 NTR , Ntrk2 , Bdnf , Trp53 , Bax , Acan , Ihh , Sox9 , Col2a1 , and Mki67 ( N = 5 per group). IF staining for ( J ) Ki-67 and ( K ) TUNEL and quantification of Ki-67 and TUNEL-positive cells in p75 +/+ and p75 −/− TMJ ( N = 5 per group). Blue, DAPI. Scale bar, 50 μm. ( L ) Cell proliferation assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). ( M ) Apoptosis assay of mBMSCs derived from p75 +/+ and p75 −/− mice ( N = 5 per group). Statistical significance was determined using unpaired two-sided Student’s t test or two-way ANOVA, as appropriate. Bonferroni correction was applied for pairwise post hoc comparisons following two-way ANOVA. Results are represented as the means ± 1 SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. N represents the number of mice or experimental replicates.

Article Snippet: For histological and morphometric analysis, pellets were cultured for 3 weeks and first imaged using a stereomicroscope (LEICA M205 FA; Leica Microsystems).

Techniques: Analysis, Staining, Gene Expression, Western Blot, TUNEL Assay, Proliferation Assay, Derivative Assay, Apoptosis Assay, Single Cell, RNA Sequencing, Real-time Polymerase Chain Reaction