Journal: Nature Communications

Article Title: Tuft cells shape airway remodeling by eliciting OXGR1- and SOX9-dependent stem cell programs

doi: 10.1038/s41467-026-70763-y

Figure Lengend Snippet: a Representative Masson’s Trichrome staining in tracheal sections at day 14. Scale bars: 200 μm. Cartilage rings are labelled C1–C6. b – e Quantification of collagen deposition over WT PBS d14 (b) or over WT ALT d14 (d), and gland numbers from C1-C7 cartilages ( c , e ). Group sizes: n = 7, 7, 5, 4, 7 ( b , c ) and n = 4, 5, 3 ( d , e ) mice, from left to right. f Experimental schema. g Representative images of lineage-labelled Sox9-tdT + cells among BCAM + BCs on the SAE at day 14. Scale bar: 200 μm. Areas outlined (a, b) are shown below (Scale bars: 50 μm). Cartilage rings are labelled C1–C7. CC Cricoid cartilage. h , i Quantification of Sox9-tdT + BCAM + cells in the SAE per high power field (HPF) ( h ) and as a percentage of total BCAM + cells ( i ) in ALT-injured trachea at day 14. Group sizes: n = 7, 15, 6 mice, from left to right. j , k Quantification of Sox9-tdT + BCAM + cells in the SAE per HPF ( j ) and as a percentage of total BCAM + cells ( k ) in influenza-injured trachea at 7 days post infection. Group sizes: n = 3, 6, 8 mice, from left to right. l , m Representative images ( l ) and quantification ( m ) of BCAM and SOX9 staining in tracheal sections at day 14. Scale bars: 20 μm. Group sizes: n = 5, 6, 4 mice, from left to right. Two-tailed unpaired Mann-Whitney U test ( b – e , m ). Kruskal–Wallis test followed by Dunn’s multiple-comparisons test with Holm–Bonferroni correction ( h – k ). The dashed outline ( g , i ) marks the border between SAE and SM. Box plots ( b – e , h – k , m ) show median (center line), 25th–75th percentiles (box), and min–max values (whiskers). Each data point represents the average of 3–4 images from one mouse, except for the glandular number ( c , e ), which represents the total glands per trachea measured by tiled imaging. See “Methods”. Panel f was created in BioRender. Lee, M. ( https://BioRender.com/dg3i7rm ). Source data are provided as a Source Data file.

Article Snippet: Female (12 weeks- to 14 weeks) and male (10 weeks- to 12 weeks) mice of the following genotypes and strains were used: C57BL/6 (Charles River Laboratories, CRL# 027), Oxgr1 -/- (KOMP, stock number 048933-UCD), Pou2f3 -/- (The Jackson Laboratory, stock number 037040), Cysltr1 -/- (kindly provided by Dr. Lora Bankova, BWH, Boston), Cysltr2 -/- (kindly provided by Dr. Joshua Boyce, BWH, Boston), Krt5 CreERT2 (The Jackson Laboratory, stock number 029155), Pou2f3 CreERT2 (The Jackson Laboratory, stock number 037511), Sox9 CreERT2 (The Jackson Laboratory, stock number 035092), Sox9 fl/fl (The Jackson Laboratory, stock number 013106), Oxgr1 fl/fl (Barrett Lab), Ltc4s fl/fl (kindly provided by Dr. Lora Bankova, BWH, Boston), and Ai9 (RCL-tdT) (The Jackson Laboratory, stock number 007909).

Techniques: Staining, Infection, Two Tailed Test, MANN-WHITNEY, Imaging

Journal: Nature Communications

Article Title: Tuft cells shape airway remodeling by eliciting OXGR1- and SOX9-dependent stem cell programs

doi: 10.1038/s41467-026-70763-y

Figure Lengend Snippet: a Representative images of DCLK1 and BCAM staining in tracheal sections from the indicated Sox9-tdT reporter strains at day 14. Sox9-tdT + BCAM + cells (white arrow). Scale bars: 50 µm. b Quantification of Sox9-tdT + BCAM + cells in the SAE from the indicated strains at day 14. Group sizes: n = 5, 6, 12, 5 mice, from left to right. Two-tailed unpaired Mann-Whitney U test. c , d Combined in situ hybridization (ISH) for Oxgr1, Krt5 and Krt8 , and immunofluorescence for SOX9 and α-SMA in tracheal sections from WT mice with or without ALT treatment ( Krt5 for BCs, Krt8 for differentiated cells, SOX9 for SMG progenitor cells, and α-SMA for SMG myoepithelial cells). Oxgr1 + SOX9 + cells (green arrows). Scale bars: 20 μm. SMG Submucosal gland. ( e ) Quantification of Oxgr1 + cells in the specified EpC subsets. Group sizes: n = 7, 16, 8, 13, 6, 10, 7, 8 mice, from left to right. Kruskal–Wallis test followed by Dunn’s multiple-comparisons test with Holm–Bonferroni correction. f Quantification of Oxgr1 + cells among the Krt5 + or Krt8 + cells. Group sizes: n = 12, 11, 12, 7 mice, from left to right. Two-tailed unpaired Mann-Whitney U test. g Experimental schema. h , i Representative images ( h ) and quantification ( i ) of BCAM and SOX9 staining in tracheal sections from the indicated strains at day 14. Scale bars: 20 μm. Group sizes: n = 5, 8, 7, 5 mice, from left to right. Kruskal–Wallis test followed by Dunn’s multiple-comparisons test with Holm–Bonferroni correction. The dashed line marks the border between SAE and SM. Box plots ( b , e , f , i ) show median (center line), 25th–75th percentiles (box), and min–max values (whiskers). Each data point represents the average of 3–4 images from one mouse. See “Methods”. Panel g was created in BioRender. Lee, M. ( https://BioRender.com/dg3i7rm ). Source data are provided as a Source Data file.

Article Snippet: Female (12 weeks- to 14 weeks) and male (10 weeks- to 12 weeks) mice of the following genotypes and strains were used: C57BL/6 (Charles River Laboratories, CRL# 027), Oxgr1 -/- (KOMP, stock number 048933-UCD), Pou2f3 -/- (The Jackson Laboratory, stock number 037040), Cysltr1 -/- (kindly provided by Dr. Lora Bankova, BWH, Boston), Cysltr2 -/- (kindly provided by Dr. Joshua Boyce, BWH, Boston), Krt5 CreERT2 (The Jackson Laboratory, stock number 029155), Pou2f3 CreERT2 (The Jackson Laboratory, stock number 037511), Sox9 CreERT2 (The Jackson Laboratory, stock number 035092), Sox9 fl/fl (The Jackson Laboratory, stock number 013106), Oxgr1 fl/fl (Barrett Lab), Ltc4s fl/fl (kindly provided by Dr. Lora Bankova, BWH, Boston), and Ai9 (RCL-tdT) (The Jackson Laboratory, stock number 007909).

Techniques: Staining, Two Tailed Test, MANN-WHITNEY, In Situ Hybridization, Immunofluorescence

Journal: Nature Communications

Article Title: Tuft cells shape airway remodeling by eliciting OXGR1- and SOX9-dependent stem cell programs

doi: 10.1038/s41467-026-70763-y

Figure Lengend Snippet: a Experimental schema. b Principal-component analysis (PCA) of the top 500 most variable genes expressed in the two groups. Numbers in parentheses indicate the percent variance captured by each PC. c Relative expression of Sox9 and canonical BC genes. d Heatmap of 1443 differentially expressed genes (DEGs) following unsupervised hierarchical clustering, defined by |log2FoldChange | > 0.58 and adjusted p-values < 0.05 with multiple testing correction using the Benjamini–Hochberg method ( e ) GSEA enrichment plots for two of the most overrepresented gene sets. Gene set enrichment analysis (GSEA) was performed using a weighted Kolmogorov–Smirnov–like statistic to calculate enrichment scores. Statistical significance was assessed by permutation testing, and nominal P values were derived from the permutation-based null distribution. Normalized enrichment scores (NES) were calculated to account for differences in gene set size, and multiple hypothesis testing was controlled using the Benjamini–Hochberg false discovery rate (FDR) correction. f – h Heatmaps of DEGs related to alarmins and inflammatory genes ( f ), collagen component genes ( g ), and ciliogenesis and cilia assembly genes ( h ). i , j Representative images ( i ) and quantification ( j ) of acetylated tubulin (Ac-tub) staining on tracheal sections from the indicated strains at day 14. Scale bars: 20 μm. The dashed line marks the border between SAE and SM. Group sizes: n = 7, 8 mice, from left to right. Two-tailed unpaired Mann-Whitney U test. Box plots ( j ) show median (center line), 25th–75th percentiles (box), and min–max values (whiskers). Each data point represents the average of 3–4 images from one mouse. See Methods. Panel a was created in BioRender. Lee, M. ( https://BioRender.com/dg3i7rm ). Source data are provided as a Source Data file.

Article Snippet: Female (12 weeks- to 14 weeks) and male (10 weeks- to 12 weeks) mice of the following genotypes and strains were used: C57BL/6 (Charles River Laboratories, CRL# 027), Oxgr1 -/- (KOMP, stock number 048933-UCD), Pou2f3 -/- (The Jackson Laboratory, stock number 037040), Cysltr1 -/- (kindly provided by Dr. Lora Bankova, BWH, Boston), Cysltr2 -/- (kindly provided by Dr. Joshua Boyce, BWH, Boston), Krt5 CreERT2 (The Jackson Laboratory, stock number 029155), Pou2f3 CreERT2 (The Jackson Laboratory, stock number 037511), Sox9 CreERT2 (The Jackson Laboratory, stock number 035092), Sox9 fl/fl (The Jackson Laboratory, stock number 013106), Oxgr1 fl/fl (Barrett Lab), Ltc4s fl/fl (kindly provided by Dr. Lora Bankova, BWH, Boston), and Ai9 (RCL-tdT) (The Jackson Laboratory, stock number 007909).

Techniques: Expressing, Derivative Assay, Staining, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Tuft cells shape airway remodeling by eliciting OXGR1- and SOX9-dependent stem cell programs

doi: 10.1038/s41467-026-70763-y

Figure Lengend Snippet: a Combined ISH for Pou2f3 , and immunofluorescence for E-Cadherin in human sinonasal tissue from healthy control (HC) and CRSwNP. Pou2f3 + cells (yellow arrow). Scale bars: 20 µm. b , c Quantification of Pou2f3 + tuft cells ( b ) and E-Cadherin staining ( c ) in the SAE across human sinonasal tissues. Group sizes: n = 6, 11, 27 independent patients, from left to right. Two-tailed unpaired Mann-Whitney U test ( b ). Kruskal–Wallis test followed by Dunn’s multiple-comparisons test with Holm–Bonferroni correction ( c ). d Spearman’s correlation analysis (two-sided) between E-Cadherin levels and Pou2f3 + tuft cell numbers in patients with CRSwNP ( n = 27, independent patients). No multiple comparison adjustment was applied. e Representative images of E-Cadherin, α-SMA, and SOX9 staining in human sinonasal mucosa from CRS patients and healthy control (HC). SOX9 + α-SMA + cells (yellow arrow). Scale bars: 20 µm. f Quantification of SOX9 + and/or α-SMA + cells in the SAE from the indicated groups. Group sizes: n = 6, 11, 27 independent patients, from left to right. Kruskal–Wallis test followed by Dunn’s multiple-comparisons test with Holm–Bonferroni correction. g , h Spearman’s correlation analysis (two-sided) between SOX9 + cell numbers and E-Cadherin intensity ( g ) and α-SMA + cell numbers ( h ) in patients with CRSwNP ( n = 27 independent patients). No multiple comparison adjustment was applied. i Spearman’s correlation analysis (two-sided) between E-Cadherin levels and SOX9 + α-SMA + cell numbers in patients with CRSwNP ( n = 27 independent patients). No multiple comparison adjustment was applied. The Spearman r values and the corresponding P values were calculated ( g – i ). j SOX9 progenitor cell signature module score in basal EpCs from the Wang scRNA-seq dataset (HRA000772) across the patients. One-sided Wilcoxon rank sum test was used. No multiple comparison adjustment was applied. The dashed line marks the border between SAE and SM. Box plots ( b , c , f ) show median (center line), 25th–75th percentiles (box), and min–max values (whiskers). Each data point represents the average of 3–4 images from one patient. See “Methods”. Source data are provided as a Source Data file.

Article Snippet: Female (12 weeks- to 14 weeks) and male (10 weeks- to 12 weeks) mice of the following genotypes and strains were used: C57BL/6 (Charles River Laboratories, CRL# 027), Oxgr1 -/- (KOMP, stock number 048933-UCD), Pou2f3 -/- (The Jackson Laboratory, stock number 037040), Cysltr1 -/- (kindly provided by Dr. Lora Bankova, BWH, Boston), Cysltr2 -/- (kindly provided by Dr. Joshua Boyce, BWH, Boston), Krt5 CreERT2 (The Jackson Laboratory, stock number 029155), Pou2f3 CreERT2 (The Jackson Laboratory, stock number 037511), Sox9 CreERT2 (The Jackson Laboratory, stock number 035092), Sox9 fl/fl (The Jackson Laboratory, stock number 013106), Oxgr1 fl/fl (Barrett Lab), Ltc4s fl/fl (kindly provided by Dr. Lora Bankova, BWH, Boston), and Ai9 (RCL-tdT) (The Jackson Laboratory, stock number 007909).

Techniques: Immunofluorescence, Control, Staining, Two Tailed Test, MANN-WHITNEY, Comparison

Journal: Nature Communications

Article Title: Tuft cells shape airway remodeling by eliciting OXGR1- and SOX9-dependent stem cell programs

doi: 10.1038/s41467-026-70763-y

Figure Lengend Snippet: (Left) In homeostasis, basal cells are the dominant progenitor of all ciliated and secretory epithelial cells on the SAE. (Right) In the setting of tissue damage elicited by allergens or by influenza ( 1 ), tuft cells generate CysLTs (2). Leukotriene E 4 (LTE 4 ) activates OXGR1 (also known as CysLTR3 and GPR99) on SOX9 + SMG progenitors ( 3 ). This results in airway remodeling including SMG hyperplasia ( 4 ) and submucosal collagen deposition ( 5 ). SOX9 + progenitors regenerate the SAE but allow persistent barrier leak ( 6 ), bringing a program that is rich in mesenchymal genes and deficient in ciliogenesis genes. This leads to an SAE with reduced ciliated cells, reduced E-Cadherin, and increased α-SMA ( 7 ). This figure was created in BioRender. Barrett, N. ( https://BioRender.com/d9cr3ur ).

Article Snippet: Female (12 weeks- to 14 weeks) and male (10 weeks- to 12 weeks) mice of the following genotypes and strains were used: C57BL/6 (Charles River Laboratories, CRL# 027), Oxgr1 -/- (KOMP, stock number 048933-UCD), Pou2f3 -/- (The Jackson Laboratory, stock number 037040), Cysltr1 -/- (kindly provided by Dr. Lora Bankova, BWH, Boston), Cysltr2 -/- (kindly provided by Dr. Joshua Boyce, BWH, Boston), Krt5 CreERT2 (The Jackson Laboratory, stock number 029155), Pou2f3 CreERT2 (The Jackson Laboratory, stock number 037511), Sox9 CreERT2 (The Jackson Laboratory, stock number 035092), Sox9 fl/fl (The Jackson Laboratory, stock number 013106), Oxgr1 fl/fl (Barrett Lab), Ltc4s fl/fl (kindly provided by Dr. Lora Bankova, BWH, Boston), and Ai9 (RCL-tdT) (The Jackson Laboratory, stock number 007909).

Techniques:

Journal: bioRxiv

Article Title: Glycogen synthase kinase 3 alpha/beta deletion induces precocious growth plate remodeling and cell loss in mice

doi: 10.1101/2020.04.04.025700

Figure Lengend Snippet: Presence of Sox9 positive cells in Gsk3a/Gsk3b KO and control mice at P36, P43 and P58 was assessed by immunohistochemistry on frontal knee joint sections (A). Bone marrow cavity-like regions are highlighted in black dotted line. Localized expression and loss of GSK3alpha (B&C) and GSK3beta (D&E) in Gsk3a/Gsk3b KO mice was evaluated by IHC at P36 (B&D) and P43 (C&E). Images are representative of 4-7 mice per group for Sox9, and 3-4 mice per group were used for GSK3alpha and GSK3beta at both time points.

Article Snippet: Primary antibodies used for immunolabelling are as follows: goat polyclonal Sox9 (AF-3075, R&D Systems), rabbit monoclonal GSK3alpha (ab40870, Abcam), rabbit polyclonal GSK3beta (12456S, Cell Signaling Technologies), mouse monoclonal type II collagen (Col2; 10R-C135b, Fitzgerald), rabbit polyclonal osteocalcin (ab93876, Abcam), rabbit polyclonal beta-catenin (bs-1165R, Bioss Antibodies) and rabbit polyclonal GLI1 (PA5-72942, Invitrogen, Thermo Fisher).

Techniques: Control, Immunohistochemistry, Expressing

Journal: BMC developmental biology

Article Title: Sonic hedgehog promotes chondrogenesis of rabbit bone marrow stem cells in a rotary cell culture system.

doi: 10.1186/s12861-019-0198-4

Figure Lengend Snippet: Fig. 5 The effect of Shh on cartilage related markers during differentiation induction in the RCCS environment. a-h RT-PCR analysis of Sox9, ACAN, collagen II, collagen X, RUNX2, ALP, PPAR-γ and leptin on days 7,14 and 21 during differentiation induction. i, j Expression of Sox 9, collagen II, ACAN, collagen X, RUNX2, ALP and PPAR-γ in the RCCS dimensional environment on days 10 and 21 of chondrogenesis. The representative results were from three independent experiments. Significant differences between the control group are indicated by * p < 0.05, **p < 0.01; differences between Shh and TGF-β transfection groups are indicated by # p < 0.05 or ## p < 0.01

Article Snippet: Antibodies recognising Ptc (Aviva Systems Biology, San Diego, CA, USA; 1:500), Smo (Aviva Systems Biology; 1:1,600), Gli1 (Biorbyt, Cambridgeshire, UK; 1:1,000), Sox9 (OriGene, Rockville, MD, USA; 1:500), collagen II (Novus Biologicals, Littleton, CO, USA; 1:200), ACAN (Novus Biologicals; 1:100), collagen X (Abcam; 1:500), RUNX2 (Abcam; 1:500), ALP (Abcam; 1:500), PPAR-γ (Santa Cruz Biotechnology; 1:1000) and GAPDH (Novus Biologicals; 1:2000) were used as primary antibodies.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Transfection

Journal: bioRxiv

Article Title: Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

doi: 10.1101/2021.11.29.470107

Figure Lengend Snippet: A-D . Heat maps show the changes in SOX9 dynamics at the single cell level per cluster during the chondrogenic differentiation of hMSCs at day 0 (A), day 2 (B), 8 (C) and 15 (D). (E) Heat map showing averaged FRAP values per cluster across time-points, letters at the right indicate cluster ID. FRAP dynamics data from three hMSC donors were combined. FRAP dynamics data from three hMSC donors were combined. To fit in the heat map scale, all the variables were normalized (0, 100). N ≥ 129. (F) Alcian blue staining show an increase in GAG production during chondrogenic differentiation of hMSCs. Arrow heads indicate the Alcian blue stained GAG at day 2, 8 and 15. Scale bar: 500 μm.

Article Snippet: The vector expressing SOX9-mGFP was constructed by cloning mGFP (PS100040, Origene) with the C-terminal of wild type SOX9 (RC208944, Origene) using SgfI and MluI restriction sites.

Techniques: Staining

Journal: bioRxiv

Article Title: Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

doi: 10.1101/2021.11.29.470107

Figure Lengend Snippet: (A-D) Heat map of FRAP rates show changes in SOX9 dynamics in undifferentiated hMSCs (A), at day 2 (B), at day 8 (C), and at day 23 (D). (E) Heat map showing averaged FRAP values per cluster across time-points, letters at the right indicate cluster ID. FRAP dynamics data from three hMSC donors were combined. To fit in the heat map scale, all the variables were normalized (0, 100). N ≥ 124. (F) ALP staining show an increase in ALP production during osteogenic differentiation of hMSCs. Black arrow heads indicate the cells with higher ALP production. Red arrow heads indicate the cells with lower ALP production. Green arrow heads indicate the cells with no ALP production. Scale bar: 500 μm.

Article Snippet: The vector expressing SOX9-mGFP was constructed by cloning mGFP (PS100040, Origene) with the C-terminal of wild type SOX9 (RC208944, Origene) using SgfI and MluI restriction sites.

Techniques: Staining

Journal: bioRxiv

Article Title: Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

doi: 10.1101/2021.11.29.470107

Figure Lengend Snippet: SOX9 dynamics in the undifferentiated hMSCs (A), day 2 (B), 8 (C) and 23 (D). (E) Heat map showing averaged FRAP values per cluster across time-points, letters at the right indicate cluster ID. FRAP dynamics data from three hMSC donors were combined. To fit in the heat map scale, all the variables were normalized (0, 100). N ≥ 125. (F) Oil Red O staining show an increase in lipid production during adipogenic differentiation of hMSCs. Black arrow heads indicate the cells with higher lipid production. Red arrow heads indicate the cells with lower lipid production. Green arrow heads indicate the cells with no lipid production. Scale bar: 500 μm.

Article Snippet: The vector expressing SOX9-mGFP was constructed by cloning mGFP (PS100040, Origene) with the C-terminal of wild type SOX9 (RC208944, Origene) using SgfI and MluI restriction sites.

Techniques: Staining

Journal: bioRxiv

Article Title: Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

doi: 10.1101/2021.11.29.470107

Figure Lengend Snippet: (A) Heat map comparing FRAP rates of SOX9-mGFP during chondro-, osteo-, and adipogenic differentiation, averaged at subpopulation level. Clusters in chondrogenic differentiation show longer Recovery half-times as compared to other differentiation lineages. Clusters in adipogenic differentiation show shorter recovery half-times as compared to other two differentiation lineages. Clusters in osteogenic differentiation show shorter recovery half-times as compared to chondrogenic and longer as compared to adipogenic differentiation lineages. Immobile fraction was higher in the initial stages chondrogenic differentiation and later stages on adipogenic differentiation. (B) Gene expression analysis by qPCR show that SOX9 and its target gene COL2A expression is increased at day 8 and 15 of chondrogenic differentiation as compared to other differentiation and time-points. During osteogenic differentiation, COL2A expression was higher than adipogenic differentiation but lower than chondrogenic differentiation (qPCR: n = 3 donors, with triplicates in each donor)

Article Snippet: The vector expressing SOX9-mGFP was constructed by cloning mGFP (PS100040, Origene) with the C-terminal of wild type SOX9 (RC208944, Origene) using SgfI and MluI restriction sites.

Techniques: Gene Expression, Expressing

Journal: bioRxiv

Article Title: Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

doi: 10.1101/2021.11.29.470107

Figure Lengend Snippet: Osteogenically differentiated hMSCs were sorted for CD10+ or CD92+ or both markers and SOX9 mobility was measured by FRAP. SOX9 mobility in FACS sorted cells show less heterogeneity. SOX9 mobility in (A) CD10+, (B) CD92+ and (D) CD10+ and CD92+ cells of osteogenically differentiated hMSCs. FRAP curves of continuous line show recovery in individual cells and symbols (square, circle and triangle) show the average. (F) Averaged FRAP curves of CD10+ and/or CD92+ are compared to osteogenically differentiated hMSCs (day 23). (E) Relationship between immobile fraction and t-half of A 2 is compared between FACS sorted (dashed lines) and Unsorted (continuous lines) cells. n ≥ 19 per condition.

Article Snippet: The vector expressing SOX9-mGFP was constructed by cloning mGFP (PS100040, Origene) with the C-terminal of wild type SOX9 (RC208944, Origene) using SgfI and MluI restriction sites.

Techniques:

Journal: bioRxiv

Article Title: Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

doi: 10.1101/2021.11.29.470107

Figure Lengend Snippet: SOX9 mobility is comparted between healthy hPCs and undifferentiated hMSCs (A and B). SOX9 mobility is comparted between healthy hPCs and chondrogenically-differentiated hMSCs at day 15 (C and D). SOX9 mobility is comparted between OA hPCs and chondrogenically-differentiated hMSCs at day 15 (C and D). n ≥ 40 per condition. hMSC control: undifferentiated hMSCs, CD: chondrogenic differentiation, D15: Day 15.

Article Snippet: The vector expressing SOX9-mGFP was constructed by cloning mGFP (PS100040, Origene) with the C-terminal of wild type SOX9 (RC208944, Origene) using SgfI and MluI restriction sites.

Techniques: Control

Journal: bioRxiv

Article Title: Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

doi: 10.1101/2021.11.29.470107

Figure Lengend Snippet: Two representative cells from each cluster of undifferentiated hMSCs are shown (A and B). Cells in cluster A exhibit discrete SOX9-mGFP localization patterns and comparatively higher expression. Cells in cluster B and C exhibit either patched or homogeneous SOX9-mGFP localization patterns. Cells in cluster D exhibit diffused SOX9-mGFP localization pattern, which results in a higher mobility with lower IF and shorter t½ of A 2 . Scale bar: 5 μm.

Article Snippet: The vector expressing SOX9-mGFP was constructed by cloning mGFP (PS100040, Origene) with the C-terminal of wild type SOX9 (RC208944, Origene) using SgfI and MluI restriction sites.

Techniques: Expressing

Journal: bioRxiv

Article Title: Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

doi: 10.1101/2021.11.29.470107

Figure Lengend Snippet: Stacked column chart at the right indicate the percentage of cells per cluster and the cluster ID is indicated at the left of the column chart. Donor 1 had considerable amount of cells in each cluster and showed moderate differentiation potential towards all lineage as compared to other donors. Donor 2 had more chondrogenic potential as compared to other two donors as evidenced by more GAG staining. This donor had 52% of cells in the cluster B, which had higher IF of SOX9. Donor 3 had more osteogenic and adipogenic potential as compared to other two donors as evidenced by more ALP and Oil Red O staining respectively. This donor had higher number of cells in the cluster B (38%) and D (36%), the later had lowest IF and shorter t-half of A 2 of SOX9. Scale bar: 500 μm.

Article Snippet: The vector expressing SOX9-mGFP was constructed by cloning mGFP (PS100040, Origene) with the C-terminal of wild type SOX9 (RC208944, Origene) using SgfI and MluI restriction sites.

Techniques: Staining

Journal: International journal of molecular sciences

Article Title: Metabolic Reconfiguration Activates Stemness and Immunomodulation of PDLSCs.

doi: 10.3390/ijms23074038

Figure Lengend Snippet: Figure 3. Curcumin increased the capabilities of osteogenic and chondrogenic differentiation in PDLSCs. (A) Heat map of RNA-seq analysis data showing the PDLSC function-related genes that were differentially regulated by curcumin treatment. (B,C) qPCR assay showed the significantly elevated levels of osteoprogenitor markers (B) and chondroprogenitor markers (C) in curcumin treated PDLSCs. (D) Alizarin red staining showed that curcumin treated PDLSCs has increased capacity to form mineralized nodules under osteoinductive conditions. (E) Western blot analysis showed the expression levels of the osteogenic genes RUNX2 and ALP were greatly increased in curcumin treated PDLSCs under osteoinductive conditions. β-actin was used as a loading control. (F) Histological analysis showed chondrogenic differentiation of vehicle and curcumin treated PDLSCs. Scale bar, 25 µm. IF staining showed increased ACAN+ and SOX9+ cells after curcumin treatment in PDLSCs. Scale bar, 25 µm. * p < 0.005. Error bars represent the s.d. from the mean values.

Article Snippet: Anti-ACAN antibody was purchased from Abcam, Cambridge, UK, and anti-SOX9 antibody was purchased from Novus Biologicals, Littleton, CO, USA.

Techniques: RNA Sequencing, Staining, Western Blot, Expressing, Control