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scrna seq library construction  (Annoroad Gene Technology Co Ltd)

 
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    Annoroad Gene Technology Co Ltd scrna seq library construction
    Scrna Seq Library Construction, supplied by Annoroad Gene Technology Co Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/scrna+seq+library+construction/pm42168200-261-8-12?v=Annoroad+Gene+Technology+Co+Ltd
    Average 86 stars, based on 1 article reviews
    scrna seq library construction - by Bioz Stars, 2026-07
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    Annoroad Gene Technology Co Ltd scrna seq library construction
    Scrna Seq Library Construction, supplied by Annoroad Gene Technology Co Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Violin plots showing the expression pattern of FOXP1/2/4 genes across different cell clusters in our previously published 8-week-old human embryonic <t>calvarial</t> <t>scRNA-seq</t> data (GSE143753). (B) Schematic illustration of the genetic lineage tracing strategy. (C) Lineage tracing strategy of Foxp4 + cell during mandible and tooth development. (D-G) IF images of E12.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (H and I) IF images of E12.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (J-M) IF images of E17.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (N and O) IF images of E16.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (P) Schematic illustration of the lineage tracing results (D-O).
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    (A) Violin plots showing the expression pattern of FOXP1/2/4 genes across different cell clusters in our previously published 8-week-old human embryonic <t>calvarial</t> <t>scRNA-seq</t> data (GSE143753). (B) Schematic illustration of the genetic lineage tracing strategy. (C) Lineage tracing strategy of Foxp4 + cell during mandible and tooth development. (D-G) IF images of E12.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (H and I) IF images of E12.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (J-M) IF images of E17.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (N and O) IF images of E16.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (P) Schematic illustration of the lineage tracing results (D-O).
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    (A) Violin plots showing the expression pattern of FOXP1/2/4 genes across different cell clusters in our previously published 8-week-old human embryonic <t>calvarial</t> <t>scRNA-seq</t> data (GSE143753). (B) Schematic illustration of the genetic lineage tracing strategy. (C) Lineage tracing strategy of Foxp4 + cell during mandible and tooth development. (D-G) IF images of E12.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (H and I) IF images of E12.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (J-M) IF images of E17.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (N and O) IF images of E16.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (P) Schematic illustration of the lineage tracing results (D-O).
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    (A) Violin plots showing the expression pattern of FOXP1/2/4 genes across different cell clusters in our previously published 8-week-old human embryonic <t>calvarial</t> <t>scRNA-seq</t> data (GSE143753). (B) Schematic illustration of the genetic lineage tracing strategy. (C) Lineage tracing strategy of Foxp4 + cell during mandible and tooth development. (D-G) IF images of E12.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (H and I) IF images of E12.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (J-M) IF images of E17.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (N and O) IF images of E16.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (P) Schematic illustration of the lineage tracing results (D-O).
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    (A) Violin plots showing the expression pattern of FOXP1/2/4 genes across different cell clusters in our previously published 8-week-old human embryonic calvarial scRNA-seq data (GSE143753). (B) Schematic illustration of the genetic lineage tracing strategy. (C) Lineage tracing strategy of Foxp4 + cell during mandible and tooth development. (D-G) IF images of E12.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (H and I) IF images of E12.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (J-M) IF images of E17.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (N and O) IF images of E16.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (P) Schematic illustration of the lineage tracing results (D-O).

    Journal: bioRxiv

    Article Title: Foxp4 + Mandibular Skeletal Stem Cells Orchestrate Bone/Tooth Development and Regeneration

    doi: 10.64898/2026.04.29.721551

    Figure Lengend Snippet: (A) Violin plots showing the expression pattern of FOXP1/2/4 genes across different cell clusters in our previously published 8-week-old human embryonic calvarial scRNA-seq data (GSE143753). (B) Schematic illustration of the genetic lineage tracing strategy. (C) Lineage tracing strategy of Foxp4 + cell during mandible and tooth development. (D-G) IF images of E12.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (H and I) IF images of E12.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E11.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (J-M) IF images of E17.5 and E19.5 mandibles in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Col2a1, Sp7 and DAPI signals were shown. The dotted circles indicated Meckel’s cartilage (MC) (n=3 mice per group from 3 independent experiments). (N and O) IF images of E16.5 and E19.5 tooth germs in Foxp4-CreERT2; tdTomato mice. Tamoxifen was administered at E16.5. The tdTomato (tdT), Emcn (blood vessel marker) and DAPI signals were shown. White dotted lines indicated the border between odontogenic epithelium and tooth germ mesenchyme. Yellow dotted lines indicated the dental follicle. mes: mesenchyme, epi: epithelium, fol: follicle (n=3 mice per group from 3 independent experiments). (P) Schematic illustration of the lineage tracing results (D-O).

    Article Snippet: For scRNA-seq library construction, Chromium Single Cell 3’ Library and Gel Bead Kit V2 (10X Genomics, PN120237) was used to generate single cell gel beads in emulsion (GEM).

    Techniques: Expressing, Marker

    (A) scRNA-seq workflow and experimental scheme. (B) UMAP visualization of 7 cell clusters in E12.5 mouse mandible. In total, 12,584 single cells from 5 embryos were analyzed. Dotted lines indicated the mdSSC cluster. (C) Violin plots showing the expression of feature genes across the 7 cell clusters. Marker genes for annotating each cell cluster were color-coded on the right. (D) Pie chart showing the cell ratios of each cell cluster. (E) UMAP visualization of the osteogenic, chondrogenic and odontogenic trajectories simulated by Monocle3. Diffusion pseudotime was indicated in the upper right panel. (F) Circle plot showing cell-cell interactions among the 7 cell clusters. The interaction strength (indicated by line thickness) was calculated by CellChat. (G) Heatmap showing regulons enriched in each cell cluster. The AUC score (row scaling) was computed. Three representative regulons (out of top 10 regulons) for each cell cluster were listed on the right. (H) Binary activities of Meis1, Sox9, Msx2 and Lhx8 regulons were shown by UMAP plots. (I) Transplantation workflow and experimental scheme. (J and K) Movat’s pentachrome staining after renal subcapsular transplantation of Foxp4 + mdSSCs (J) or control (K) cells for 4 weeks. Cartilage (i) and bone (ii) formation were shown. The tdTomato (tdT) signal indicated donor-derived cells (J, lower left panel). (L) Quantification of the renal subcapsular transplantation data (n=10 mice per group from 5 independent experiments). (M) Representative microCT image of the ectopic tooth formed by Foxp4 + mdSSCs 4 weeks after nasal sinus transplantation. Arrowheads indicated the tooth cups. (N and O) Safranin O/Fast Green (N) and IF (O) staining of the ectopic tooth formed by Foxp4 + mdSSCs 4 weeks after nasal sinus transplantation. Foxp4 + mdSSCs from E12.5 Foxp4-CreERT2; tdTomato embryos were transplanted. ame: ameloblast, ena: enamel, den: dentin, od: odontoblast. (P and Q) Safranin O/Fast Green (P) and IF (Q) staining of the native tooth in the recipient mice. ame: ameloblast, ena: enamel, den: dentin, od: odontoblast.

    Journal: bioRxiv

    Article Title: Foxp4 + Mandibular Skeletal Stem Cells Orchestrate Bone/Tooth Development and Regeneration

    doi: 10.64898/2026.04.29.721551

    Figure Lengend Snippet: (A) scRNA-seq workflow and experimental scheme. (B) UMAP visualization of 7 cell clusters in E12.5 mouse mandible. In total, 12,584 single cells from 5 embryos were analyzed. Dotted lines indicated the mdSSC cluster. (C) Violin plots showing the expression of feature genes across the 7 cell clusters. Marker genes for annotating each cell cluster were color-coded on the right. (D) Pie chart showing the cell ratios of each cell cluster. (E) UMAP visualization of the osteogenic, chondrogenic and odontogenic trajectories simulated by Monocle3. Diffusion pseudotime was indicated in the upper right panel. (F) Circle plot showing cell-cell interactions among the 7 cell clusters. The interaction strength (indicated by line thickness) was calculated by CellChat. (G) Heatmap showing regulons enriched in each cell cluster. The AUC score (row scaling) was computed. Three representative regulons (out of top 10 regulons) for each cell cluster were listed on the right. (H) Binary activities of Meis1, Sox9, Msx2 and Lhx8 regulons were shown by UMAP plots. (I) Transplantation workflow and experimental scheme. (J and K) Movat’s pentachrome staining after renal subcapsular transplantation of Foxp4 + mdSSCs (J) or control (K) cells for 4 weeks. Cartilage (i) and bone (ii) formation were shown. The tdTomato (tdT) signal indicated donor-derived cells (J, lower left panel). (L) Quantification of the renal subcapsular transplantation data (n=10 mice per group from 5 independent experiments). (M) Representative microCT image of the ectopic tooth formed by Foxp4 + mdSSCs 4 weeks after nasal sinus transplantation. Arrowheads indicated the tooth cups. (N and O) Safranin O/Fast Green (N) and IF (O) staining of the ectopic tooth formed by Foxp4 + mdSSCs 4 weeks after nasal sinus transplantation. Foxp4 + mdSSCs from E12.5 Foxp4-CreERT2; tdTomato embryos were transplanted. ame: ameloblast, ena: enamel, den: dentin, od: odontoblast. (P and Q) Safranin O/Fast Green (P) and IF (Q) staining of the native tooth in the recipient mice. ame: ameloblast, ena: enamel, den: dentin, od: odontoblast.

    Article Snippet: For scRNA-seq library construction, Chromium Single Cell 3’ Library and Gel Bead Kit V2 (10X Genomics, PN120237) was used to generate single cell gel beads in emulsion (GEM).

    Techniques: Expressing, Marker, Diffusion-based Assay, Transplantation Assay, Staining, Control, Derivative Assay

    (A) scRNA-seq workflow and experimental scheme. (B) UMAP visualization of 5 cell clusters in 12-week-old adult mandible. In total, 11,862 single cells from 6 mice were analyzed. Dotted lines indicated the mdSSC cluster. (C) Violin plots showing the expression of feature genes across the 5 cell clusters. Marker genes for annotating each cell cluster were color-coded on the right. (D) Pie chart showing the cell ratios of each cell cluster. (E) UMAP visualization of the osteogenesis and fibrogenesis trajectories simulated by Monocle3. Diffusion pseudotime was indicated in the upper right panel. (F) Circle plot showing cell-cell interactions among the 5 cell clusters. The interaction strength (indicated by line thickness) was calculated by CellChat. (G) Heatmap showing regulons enriched in each cell cluster. The AUC score (row scaling) was computed. Three representative regulons (out of top 10 regulons) for each cell cluster were listed on the right. (H) Binary activities of Sox9, Sp7, Npdc1 and Trp63 regulons were shown by UMAP plots. (I and J) IF images of Foxp4 + mdSSCs in adult mandibular (I) and alveolar (J) bone. Tamoxifen was administered for 5 consecutive days in 12-week-old Foxp4-CreERT2; tdTomato mice and analyzed 1 day after. The tdTomato (tdT), Acan and DAPI signals were shown. Dotted lines (J) indicated the border of the dental crown (n = 3 mice per group from 3 independent experiments). Arrowheads indicated the tdT + Acan + mdSSCs. md: mandibular bone, alv: alveolar bone. (K) Movat’s pentachrome staining showing cartilage (i) and bone (ii) formation after renal subcapsular transplantation of adult Foxp4 + mdSSCs from 12-week-old Foxp4-CreERT2; tdTomato mice. The tdTomato (tdT) signal (lower left panel) demonstrated donor-derived cells. (L) Quantification of the renal subcapsular transplantation data (n=10 mice per group from 8 independent experiments). (M) Representative microCT image of the ectopic tooth formed by adult Foxp4 + mdSSCs 4 weeks after nasal sinus transplantation. Arrowheads indicated the tooth cups. (N and O) Safranin O/Fast Green (N) and IF (O) staining of the ectopic tooth formed by adult Foxp4 + mdSSCs 4 weeks after nasal sinus transplantation. Foxp4 + mdSSCs from 12-week-old Foxp4-CreERT2; tdTomato mice were transplanted.

    Journal: bioRxiv

    Article Title: Foxp4 + Mandibular Skeletal Stem Cells Orchestrate Bone/Tooth Development and Regeneration

    doi: 10.64898/2026.04.29.721551

    Figure Lengend Snippet: (A) scRNA-seq workflow and experimental scheme. (B) UMAP visualization of 5 cell clusters in 12-week-old adult mandible. In total, 11,862 single cells from 6 mice were analyzed. Dotted lines indicated the mdSSC cluster. (C) Violin plots showing the expression of feature genes across the 5 cell clusters. Marker genes for annotating each cell cluster were color-coded on the right. (D) Pie chart showing the cell ratios of each cell cluster. (E) UMAP visualization of the osteogenesis and fibrogenesis trajectories simulated by Monocle3. Diffusion pseudotime was indicated in the upper right panel. (F) Circle plot showing cell-cell interactions among the 5 cell clusters. The interaction strength (indicated by line thickness) was calculated by CellChat. (G) Heatmap showing regulons enriched in each cell cluster. The AUC score (row scaling) was computed. Three representative regulons (out of top 10 regulons) for each cell cluster were listed on the right. (H) Binary activities of Sox9, Sp7, Npdc1 and Trp63 regulons were shown by UMAP plots. (I and J) IF images of Foxp4 + mdSSCs in adult mandibular (I) and alveolar (J) bone. Tamoxifen was administered for 5 consecutive days in 12-week-old Foxp4-CreERT2; tdTomato mice and analyzed 1 day after. The tdTomato (tdT), Acan and DAPI signals were shown. Dotted lines (J) indicated the border of the dental crown (n = 3 mice per group from 3 independent experiments). Arrowheads indicated the tdT + Acan + mdSSCs. md: mandibular bone, alv: alveolar bone. (K) Movat’s pentachrome staining showing cartilage (i) and bone (ii) formation after renal subcapsular transplantation of adult Foxp4 + mdSSCs from 12-week-old Foxp4-CreERT2; tdTomato mice. The tdTomato (tdT) signal (lower left panel) demonstrated donor-derived cells. (L) Quantification of the renal subcapsular transplantation data (n=10 mice per group from 8 independent experiments). (M) Representative microCT image of the ectopic tooth formed by adult Foxp4 + mdSSCs 4 weeks after nasal sinus transplantation. Arrowheads indicated the tooth cups. (N and O) Safranin O/Fast Green (N) and IF (O) staining of the ectopic tooth formed by adult Foxp4 + mdSSCs 4 weeks after nasal sinus transplantation. Foxp4 + mdSSCs from 12-week-old Foxp4-CreERT2; tdTomato mice were transplanted.

    Article Snippet: For scRNA-seq library construction, Chromium Single Cell 3’ Library and Gel Bead Kit V2 (10X Genomics, PN120237) was used to generate single cell gel beads in emulsion (GEM).

    Techniques: Expressing, Marker, Diffusion-based Assay, Staining, Transplantation Assay, Derivative Assay

    (A-D) Dot plots showing GO terms and KEGG pathways enriched in the mdSSC subset of E12.5 and 12-week-old mandible scRNA-seq data. BP: Biological process. (E) Bulk RNA-seq workflow and experimental scheme. Tamoxifen was administered for 1 day at E11.5. Mouse mandibles were analyzed at E12.5. (F) Dot plot showing GO terms enriched in Foxp4+ SSCs (mdSSC) as compared to Foxp4-SSCs by analyzing E12.5 bulk RNA-seq data (E). BP: Biological process. (G) Box plot showing relative enrichment of intraflagellar transport genes in Foxp4+ SSCs (mdSSC) as compared to Foxp4- SSCs by analyzing E12.5 bulk RNA-seq data (E). (H) qPCR analysis of Foxp4+ mdSSCs following Ift140 or control siRNA knockdown (n=3 independent experiments). (I) Crystal violet staining of CFU-F colonies with quantification (n=3 independent experiments). (J-L) In vitro trilineage differentiation. Alizarin red (J), Oil Red O (K) and Toluidine blue (L) staining showing osteogenic, adipogenic and chondrogenic differentiation, respectively. qPCR analyses of adipogenic, osteogenic and chondrogenic markers were shown (n=3 independent experiments). Statistical significance was determined by unpaired two-tailed Student’s t test. All data represent mean ± SD (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001).

    Journal: bioRxiv

    Article Title: Foxp4 + Mandibular Skeletal Stem Cells Orchestrate Bone/Tooth Development and Regeneration

    doi: 10.64898/2026.04.29.721551

    Figure Lengend Snippet: (A-D) Dot plots showing GO terms and KEGG pathways enriched in the mdSSC subset of E12.5 and 12-week-old mandible scRNA-seq data. BP: Biological process. (E) Bulk RNA-seq workflow and experimental scheme. Tamoxifen was administered for 1 day at E11.5. Mouse mandibles were analyzed at E12.5. (F) Dot plot showing GO terms enriched in Foxp4+ SSCs (mdSSC) as compared to Foxp4-SSCs by analyzing E12.5 bulk RNA-seq data (E). BP: Biological process. (G) Box plot showing relative enrichment of intraflagellar transport genes in Foxp4+ SSCs (mdSSC) as compared to Foxp4- SSCs by analyzing E12.5 bulk RNA-seq data (E). (H) qPCR analysis of Foxp4+ mdSSCs following Ift140 or control siRNA knockdown (n=3 independent experiments). (I) Crystal violet staining of CFU-F colonies with quantification (n=3 independent experiments). (J-L) In vitro trilineage differentiation. Alizarin red (J), Oil Red O (K) and Toluidine blue (L) staining showing osteogenic, adipogenic and chondrogenic differentiation, respectively. qPCR analyses of adipogenic, osteogenic and chondrogenic markers were shown (n=3 independent experiments). Statistical significance was determined by unpaired two-tailed Student’s t test. All data represent mean ± SD (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001).

    Article Snippet: For scRNA-seq library construction, Chromium Single Cell 3’ Library and Gel Bead Kit V2 (10X Genomics, PN120237) was used to generate single cell gel beads in emulsion (GEM).

    Techniques: RNA Sequencing, Control, Knockdown, Staining, In Vitro, Two Tailed Test

    (A) scRNA-seq workflow and experimental scheme. The mandibular bone fragment obtained from a 29-year-old patient was analyzed. (B) UMAP visualization of the 6 cell clusters in a previously published human mandible scRNA-seq data (GSE289393). In total, 9,327 cells were analyzed. Dotted lines indicated the mdSSC cluster. (C) Violin plots showing the expression of feature genes across the 6 cell clusters. Marker genes for annotating each cell cluster were color-coded on the right. (D) Heatmap showing the regulons enriched in each cell cluster. The AUC score (row scaling) was computed. Three representative regulons (out of top 10 regulons) for each cell cluster were listed on the right. (E) Binary activities of FOXP4, SOX9, GLI3 and TWIST1 regulons were shown by UMAP plots. (F and G) Dot plots showing GO and KEGG terms enriched in the mdSSC subset. BP: Biological process. (H) Transplantation workflow and experimental scheme. The mandibular bone fragments obtained from 24 to 29-year-old patients were analyzed. (I) Movat’s pentachrome staining showing cartilage (i) and bone (ii) formation after renal subcapsular transplantation of human mdSSCs. Lower left panel: IF images of human Vimentin (hVimentin) to demonstrate donor-derived cells. (J) Quantification of the renal subcapsular transplantation data (n=10 or 16 recipient mice per group from 5 independent experiments). (K) Representative microCT image of the ectopic tooth formed by human mdSSCs 4 weeks after nasal sinus transplantation into NCG mice. Arrowheads indicated the tooth cups. (L and M) Safranin O/Fast Green (L) and IF (M) staining of the ectopic tooth formed by human mdSSCs 4 weeks after nasal sinus transplantation into NCG mice. ame: ameloblast, den: dentin, od: odontoblast, epi: epithelium. Human Vimentin, DSPP and DAPI signals were shown in (M). (N) Representative microCT image of the ectopic tooth formed by human mdSSCs 8 weeks after nasal sinus transplantation into NCG mice. Arrowheads indicated the tooth cups. Dotted line indicated the boundary between dental crown and root. Dotted box indicated the root apex. (O) Safranin O/Fast Green staining of the ectopic tooth formed by human mdSSCs 8 weeks after nasal sinus transplantation into NCG mice. den: dentin, od: odontoblast. Arrowhead indicated the root apex. (P) Representative microCT image of a 14-year-old human premolar. Arrowheads indicated the tooth cups. Dotted lines indicated boundary between dental crown and root. Dotted box indicated the root apex. (Q) Safranin O/Fast Green staining of the 14-year-old human premolar. Arrowhead indicated the root apex. den: dentin, od: odontoblast.

    Journal: bioRxiv

    Article Title: Foxp4 + Mandibular Skeletal Stem Cells Orchestrate Bone/Tooth Development and Regeneration

    doi: 10.64898/2026.04.29.721551

    Figure Lengend Snippet: (A) scRNA-seq workflow and experimental scheme. The mandibular bone fragment obtained from a 29-year-old patient was analyzed. (B) UMAP visualization of the 6 cell clusters in a previously published human mandible scRNA-seq data (GSE289393). In total, 9,327 cells were analyzed. Dotted lines indicated the mdSSC cluster. (C) Violin plots showing the expression of feature genes across the 6 cell clusters. Marker genes for annotating each cell cluster were color-coded on the right. (D) Heatmap showing the regulons enriched in each cell cluster. The AUC score (row scaling) was computed. Three representative regulons (out of top 10 regulons) for each cell cluster were listed on the right. (E) Binary activities of FOXP4, SOX9, GLI3 and TWIST1 regulons were shown by UMAP plots. (F and G) Dot plots showing GO and KEGG terms enriched in the mdSSC subset. BP: Biological process. (H) Transplantation workflow and experimental scheme. The mandibular bone fragments obtained from 24 to 29-year-old patients were analyzed. (I) Movat’s pentachrome staining showing cartilage (i) and bone (ii) formation after renal subcapsular transplantation of human mdSSCs. Lower left panel: IF images of human Vimentin (hVimentin) to demonstrate donor-derived cells. (J) Quantification of the renal subcapsular transplantation data (n=10 or 16 recipient mice per group from 5 independent experiments). (K) Representative microCT image of the ectopic tooth formed by human mdSSCs 4 weeks after nasal sinus transplantation into NCG mice. Arrowheads indicated the tooth cups. (L and M) Safranin O/Fast Green (L) and IF (M) staining of the ectopic tooth formed by human mdSSCs 4 weeks after nasal sinus transplantation into NCG mice. ame: ameloblast, den: dentin, od: odontoblast, epi: epithelium. Human Vimentin, DSPP and DAPI signals were shown in (M). (N) Representative microCT image of the ectopic tooth formed by human mdSSCs 8 weeks after nasal sinus transplantation into NCG mice. Arrowheads indicated the tooth cups. Dotted line indicated the boundary between dental crown and root. Dotted box indicated the root apex. (O) Safranin O/Fast Green staining of the ectopic tooth formed by human mdSSCs 8 weeks after nasal sinus transplantation into NCG mice. den: dentin, od: odontoblast. Arrowhead indicated the root apex. (P) Representative microCT image of a 14-year-old human premolar. Arrowheads indicated the tooth cups. Dotted lines indicated boundary between dental crown and root. Dotted box indicated the root apex. (Q) Safranin O/Fast Green staining of the 14-year-old human premolar. Arrowhead indicated the root apex. den: dentin, od: odontoblast.

    Article Snippet: For scRNA-seq library construction, Chromium Single Cell 3’ Library and Gel Bead Kit V2 (10X Genomics, PN120237) was used to generate single cell gel beads in emulsion (GEM).

    Techniques: Expressing, Marker, Transplantation Assay, Staining, Derivative Assay