Journal: bioRxiv

Article Title: Pharmacodynamic and stage-dependent therapeutic efficacy of SFRP1 neutralization in a mouse model of Alzheimer’s disease

doi: 10.64898/2026.03.26.714543

Figure Lengend Snippet: ELISA quantification of biotinylated α-SFRP1 mAb levels in (A) serum and (B) brain RIPA-soluble homogenates from 4-8 month-old APP/PS1 mice. Samples were collected 6 and 24 h after a single 100 µg intraperitoneal or retro-orbital injection of α-SFRP1. “Pre-Inj.” Indicates mAb levels before the injection (expected to be zero). Data are presented as mean ± SEM. Statistical analyses were performed using (A) two-way ANOVA followed by Sidak’s post hoc test for multiple comparisons or (B) Mann-Whitney test.

Article Snippet: Mice divided in different groups and indicated in each specific case, were treated with either α-SFRP1 mAb or a non-specific IgG1 mAb as control (IgG1, MAB002, Bio-Techne, MN, USA), or with SFRP1 inhibitor WAY-316606 (WAY; S5815, Selleck Chemicals, USA) using, in this case, the DMSO diluent as control.

Techniques: Enzyme-linked Immunosorbent Assay, Injection, MANN-WHITNEY

Journal: bioRxiv

Article Title: Pharmacodynamic and stage-dependent therapeutic efficacy of SFRP1 neutralization in a mouse model of Alzheimer’s disease

doi: 10.64898/2026.03.26.714543

Figure Lengend Snippet: (A) Representative transverse whole-body PET-MRI maximum-intensity projection images acquired at 2, 10, 24, and 168 h after tail-vein administration of 89 Zr-IgG1 (left) or 89 Zr-α-SFRP1.10 (right). (B-G) In vivo quantification of 89 Zr-IgG1 and 89 Zr-α-SFRP1 overtime in (B) brain, (C) liver, (D) kidneys, (E) lungs, (F) heart, (G) and bladder. Data are presented as standardized uptake values (SUV) and shown as mean ± SEM. Statistical analysis were performed using two-way ANOVA followed by Sidak’s post-hoc correction for multiple comparisons.

Article Snippet: Mice divided in different groups and indicated in each specific case, were treated with either α-SFRP1 mAb or a non-specific IgG1 mAb as control (IgG1, MAB002, Bio-Techne, MN, USA), or with SFRP1 inhibitor WAY-316606 (WAY; S5815, Selleck Chemicals, USA) using, in this case, the DMSO diluent as control.

Techniques: In Vivo

Journal: bioRxiv

Article Title: Pharmacodynamic and stage-dependent therapeutic efficacy of SFRP1 neutralization in a mouse model of Alzheimer’s disease

doi: 10.64898/2026.03.26.714543

Figure Lengend Snippet: (A) Representative transversal brain PET-MRI maximum-intensity projection images acquired at 2, 10, 24, and 168 h after intravenous tail administration of 89 Zr-IgG1 (bottom) or 89 Zr-α-SFRP1 (top). The left-most image corresponds to the mouse brain template used for the analysis. Volumes of interest include the olfactory bulb (purple), cerebral cortex (green), striatum (orange), hippocampus (blue), thalamus (red), and cerebellum (yellow). (B, C) In vivo quantification of 89 Zr-IgG1 and 89 Zr-α-SFRP1 over time in the brain regions of primary interest: (B) cortex and (C) hippocampus. Data are presented as standardized uptake values (SUV) and shown as mean ± SEM. Statistical analysis were performed using two-way ANOVA followed by Sidak’s post-hoc correction for multiple comparisons.

Article Snippet: Mice divided in different groups and indicated in each specific case, were treated with either α-SFRP1 mAb or a non-specific IgG1 mAb as control (IgG1, MAB002, Bio-Techne, MN, USA), or with SFRP1 inhibitor WAY-316606 (WAY; S5815, Selleck Chemicals, USA) using, in this case, the DMSO diluent as control.

Techniques: Olfactory, In Vivo

Journal: bioRxiv

Article Title: Pharmacodynamic and stage-dependent therapeutic efficacy of SFRP1 neutralization in a mouse model of Alzheimer’s disease

doi: 10.64898/2026.03.26.714543

Figure Lengend Snippet: ( A ) Schematic of the experimental design. 4-month-old homozygous APP/PS1 mice received weekly retro-orbital injections of IgG1 or α-SFRP1.10 (100 µg) for 5 months; analyses in panels ( B–G ) were performed at 9 months of age. The time diagram indicates the expected age at which these mice initiate amyloid-β plaque accumulation. Illustration from NIAID NIH BioArt Source ( https://bioart.niaid.nih.gov/ ). ( B ) ELISA quantification of SFRP1 levels in brain RIPA-soluble homogenates. ( C ) Representative confocal images of cortices (top) and hippocampi (bottom) from IgG1-treated (left) and α-SFRP1.10-treated (right) mice stained with Thioflavin S (ThioS) to label amyloid plaque cores. Scale bar: 100 µm. Quantification of the number (top) and mean plaque area (bottom) of ( D ) ThioS+ plaques and ( E ) LAMP1-positive puncta in the cortex (left) and hippocampus (right). Quantification of (F) GFAP and (G) Iba1 fluorescence intensity in the cortex (left) and hippocampus (right), expressed as fold change relative to the mean value of the IgG1 group. Quantification analyses were performed for individual mice, averaging the results of 3-8 slices per mouse. Data are presented as mean ± SEM. Statistical analyses were performed using Student’s t test.

Article Snippet: Mice divided in different groups and indicated in each specific case, were treated with either α-SFRP1 mAb or a non-specific IgG1 mAb as control (IgG1, MAB002, Bio-Techne, MN, USA), or with SFRP1 inhibitor WAY-316606 (WAY; S5815, Selleck Chemicals, USA) using, in this case, the DMSO diluent as control.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Fluorescence

Journal: bioRxiv

Article Title: Pharmacodynamic and stage-dependent therapeutic efficacy of SFRP1 neutralization in a mouse model of Alzheimer’s disease

doi: 10.64898/2026.03.26.714543

Figure Lengend Snippet: ( A ) Schematic of the experimental design. 4-month-old homozygous APP/PS1 mice received weekly retro-orbital injections of IgG1 or α-SFRP1.10 (200 µg) for 2 months; analyses in panels ( B–H ) were performed at 6 months of age. The time diagram indicates the expected age at which these mice initiate amyloid-β plaque accumulation. Illustration from NIAID NIH BioArt Source ( https://bioart.niaid.nih.gov/ ). ( B ) Kaplan–Meier curves showing mouse survival during the treatment period; each vertical step represents a death event. ( C ) ELISA quantification of SFRP1 levels in brain RIPA-soluble homogenates. ( D ) Representative confocal images of cortices (top) and hippocampi (bottom) from IgG1-treated (left) and α-SFRP1.10-treated (right) mice stained with Thioflavin S (ThioS) to label amyloid plaque cores. Scale bar: 100 µm. Quantification of the number (top) and mean plaque area (bottom) of ( E ) ThioS+ plaques and ( F ) LAMP1+ puncta in the cortex (left) and hippocampus (right). Quantification of (G) GFAP and (H) Iba1 fluorescence intensity in the cortex (left) and hippocampus (right), expressed as fold change relative to the mean value of the IgG1 group. Quantification analyses were performed for individual mice, averaging the results of 3-8 slices per mouse. Data are presented as mean ± SEM. Statistical analyses were performed using Student’s t test.

Article Snippet: Mice divided in different groups and indicated in each specific case, were treated with either α-SFRP1 mAb or a non-specific IgG1 mAb as control (IgG1, MAB002, Bio-Techne, MN, USA), or with SFRP1 inhibitor WAY-316606 (WAY; S5815, Selleck Chemicals, USA) using, in this case, the DMSO diluent as control.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Fluorescence

Journal: bioRxiv

Article Title: Pharmacodynamic and stage-dependent therapeutic efficacy of SFRP1 neutralization in a mouse model of Alzheimer’s disease

doi: 10.64898/2026.03.26.714543

Figure Lengend Snippet: ( A ) Schematic of the experimental design. 8-month-old heterozygous APP/PS1 mice received weekly retro-orbital injections of WAY-316606 (1 µM) or vehicle (2% DMSO) for 2 months; analyses in panels ( B–D ) were performed at 10 months of age. The time diagram indicates the expected age at which these mice initiate amyloid-β plaque accumulation. Illustration from NIAID NIH BioArt Source ( https://bioart.niaid.nih.gov/ ). ( B ) ELISA quantification of SFRP1 levels in brain RIPA-soluble homogenates. ( C ) Representative confocal images of cortices (top) and hippocampi (bottom) from vehicle-treated (left), WAY-316606-treated (center) mice stained with ThioS to label amyloid plaque cores. Scale bar: 100 µm. Quantification of number (top) and mean plaque area (bottom) of ( D ) ThioS+ plaque and LAMP1+ puncta ( E ) in the cortex (left) and hippocampus (right). Quantification analyses were performed for individual mice, averaging the results of 3-8 slices per mouse. Data are presented as mean ± SEM. Statistical analyses were performed using Student’s t test.

Article Snippet: Mice divided in different groups and indicated in each specific case, were treated with either α-SFRP1 mAb or a non-specific IgG1 mAb as control (IgG1, MAB002, Bio-Techne, MN, USA), or with SFRP1 inhibitor WAY-316606 (WAY; S5815, Selleck Chemicals, USA) using, in this case, the DMSO diluent as control.

Techniques: Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Orthopaedic Translation

Article Title: Silencing SFRP1 in bone mesenchymal stem cells alleviates pediatric B-ALL-driven bone loss by activating Wnt/β-catenin signaling

doi: 10.1016/j.jot.2026.101071

Figure Lengend Snippet: SFRP1 suppresses osteogenesis of B-ALL–derived BMSCs through the Wnt/β-catenin pathway. (A) Venn diagram showing overlap between upregulated mRNA and protein datasets. (B) Protein-protein interaction network of DEPs (red: upregulated; green: downregulated). (C) KEGG pathway enrichment analysis of DEGs. (D, E) Quantification of SFRP1 protein expression in hBMSCs from hCTR and hB-ALL patients. (F, G) Quantification of osteogenic proteins (COL1A1, ALP, RUNX2) in hBMSCs from hB-ALL with WAY-316606. (H, I) Quantification of SFRP1 protein expression in mBMSCs from mCTR and mB-ALL mice. (J, M) β-catenin and RUNX2 protein expression in mBMSCs with different treatments. (K, L) Immunofluorescence imaging and quantification of SFRP1 (red) in mouse trabecular bone. (N–P) Immunohistochemical analysis of β-catenin and RUNX2 expression in mouse trabecular bone (n = 3; ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001).

Article Snippet: Paraffin-embedded femoral sections underwent antigen retrieval, blocking with 3% BSA, and incubation with anti-SFRP1 antibody (Proteintech, 26460-1-AP, 1:200) for 12 h, followed by Alexa Fluor 594-conjugated secondary antibody (Abcam, ab150080, 1:500).

Techniques: Derivative Assay, Expressing, Immunofluorescence, Imaging, Immunohistochemical staining

Journal: Journal of Orthopaedic Translation

Article Title: Silencing SFRP1 in bone mesenchymal stem cells alleviates pediatric B-ALL-driven bone loss by activating Wnt/β-catenin signaling

doi: 10.1016/j.jot.2026.101071

Figure Lengend Snippet: SFRP1 inhibition enhances β-catenin expression and nuclear translocation (A, C) Immunofluorescence analysis and quantification of SFRP1 expression (red) in femoral sections of mB-ALL mice across treatment groups. (B, D) β-catenin expression and nuclear localization in femoral trabeculae across treatment groups (n = 5; ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001).

Article Snippet: Paraffin-embedded femoral sections underwent antigen retrieval, blocking with 3% BSA, and incubation with anti-SFRP1 antibody (Proteintech, 26460-1-AP, 1:200) for 12 h, followed by Alexa Fluor 594-conjugated secondary antibody (Abcam, ab150080, 1:500).

Techniques: Inhibition, Expressing, Translocation Assay, Immunofluorescence

Journal: Journal of Orthopaedic Translation

Article Title: Silencing SFRP1 in bone mesenchymal stem cells alleviates pediatric B-ALL-driven bone loss by activating Wnt/β-catenin signaling

doi: 10.1016/j.jot.2026.101071

Figure Lengend Snippet: shSFRP1@Lipo-E7 targets BMSCs to regulate the SFRP1/β-catenin signaling pathway, promoting osteogenic differentiation of BMSCs and alleviating bone loss induced by B-ALL. (A) B-ALL cells induce abnormal elevation of SFRP1 in BMSCs, promoting adipogenic differentiation and disrupting the osteogenic-osteoclastic coupling balance, ultimately leading to bone loss. (B) Tail vein delivery of shSFRP1-loaded BMSC-targeted liposomes downregulates SFRP1 to enhance BMSC osteogenic differentiation, suppress adipogenic differentiation, and indirectly inhibit osteoclastogenesis by remodeling the bone microenvironment. (C) SFRP1 inhibit Wnt signaling by promoting the phosphorylation of β-catenin and reducing its nuclear translocation.

Article Snippet: Paraffin-embedded femoral sections underwent antigen retrieval, blocking with 3% BSA, and incubation with anti-SFRP1 antibody (Proteintech, 26460-1-AP, 1:200) for 12 h, followed by Alexa Fluor 594-conjugated secondary antibody (Abcam, ab150080, 1:500).

Techniques: Liposomes, Phospho-proteomics, Translocation Assay

Journal: Nature Communications

Article Title: Spatiotemporal interplay between epithelial and mesenchymal cells drives human dentinogenesis

doi: 10.1038/s41467-026-69545-3

Figure Lengend Snippet: a Split violin plots showing the expression distribution of WNT and NOTCH pathway components in DE (red) and DPL (blue). Each dot represents a single cell. Adjacent pie charts quantify the percentage of cells within the DE population expressing specific WNT and NOTCH ligands. b Dot plots illustrating the dynamic expression of WNT and NOTCH pathway genes in DE (left panel, red color scale) and DPL (right panel, blue color scale) across developmental stages. Dot size is proportional to the percentage of cells expressing the gene, and color intensity reflects the average z-score normalized expression level. PCW, post-conception weeks; Y, years. c Line graphs tracking the average expression of key WNT ( WNT6 , WNT7B , WNT10B ) and NOTCH ( JAG1 ) ligands within the DE population over time. d IF validation of the spatial localization of key WNT and NOTCH pathway proteins across different developmental stages. Target proteins (WNT6, WNT7B, WNT10B, DKK1, SFRP1, LRP6, LEF1, JAG1, NOTCH2, HEY1) are shown in green or red, with nuclei counterstained with DAPI (blue). Dotted lines outline the epithelial-mesenchymal boundary. Insets show magnified views of the indicated regions. Images are representative of experiments that were independently repeated three times with similar results ( n = 3 biological replicates). PT permanent tooth. Scale bars: 100 μm. Data in ( a – c ) are derived from scRNA-seq analysis of n = 2 biological replicates per developmental stage.

Article Snippet: Primary antibodies were applied against: DSPP (Abcam, ab216892), Ki67 (Cell Signaling Technology, 9129, Clone D3B5), KRT14 (Abcam, ab119695, Clone LL002), WNT6 (Abcam, ab50030), WNT7B (Proteintech, 10605-1-AP), WNT10B (Abcam, ab70816), LRP6 (Santa Cruz, sc-25317, Clone C-10), JAG1 (Abcam, ab300561, Clone EPR26388-51), NOTCH2 (Abcam, ab313453, Clone EPR28701-38), DKK1 (Abcam, ab61034, Clone EPR4759), SFRP1 (Proteintech, 26460-1-AP), HEY1 (Proteintech, 19929-1-AP), LEF1 (Abcam, ab137872, Clone EPR2029Y), BMP2 (Abcam,ab214821), BMP4 (Abcam, ab39973), GFP(CST, 2956, Clone D5.1) and rabbit IgG isotype control (CST, 3900, Clone DA1E) were added.

Techniques: Expressing, Single Cell, Biomarker Discovery, Derivative Assay

Journal: Nature Communications

Article Title: Spatiotemporal interplay between epithelial and mesenchymal cells drives human dentinogenesis

doi: 10.1038/s41467-026-69545-3

Figure Lengend Snippet: a Vector field visualization (left) and heatmap of branch-dependent differentially expressed genes (DEGs) (right) along differentiation trajectories. Branch-dependent DEGs were identified using Branched Expression Analysis Modeling (q-value < 0.0001). b UMAP visualization of DPL subpopulations (right) with temporal distribution of cell proportions (left). c Expression distribution of genes across identified clusters. d RNAscope staining of DLX6-AS1 (red, arrowheads) in the human molar. Nuclei are DAPI-stained (blue), and dotted lines mark the epithelial-mesenchymal boundary. Images are representative of experiments independently repeated three times with similar results ( n = 3 biological replicates). Scale bars: 100 μm for molars at 10 and 12 PCW; 200 μm for molars at 24 PCW and 13 Y. e Volcano plot comparing DLX6-AS1 ⁺ and NR2F1 ⁺ TWIST1 ⁺ DP (red: DLX6-AS1 ⁺ DP-enriched; blue: NR2F1 + TWIST1 + DP-enriched; gray: non-significant). Dashed lines mark significance thresholds (adjusted P < 0.05, |log₂FC | > 1.0), determined by a two-sided Wilcoxon rank-sum test with Bonferroni correction. Genes with adjusted p values of zero are shown as upward-pointing arrows at the top of the y-axis to indicate that their significance exceeds the plotted range. f Spatial distribution (top) and UMAP FeaturePlots (bottom) of DLX6-AS1 , FZD2 , LEF1 , NOTCH2 , and HEY1 . g Temporal transcriptional dynamics in DLX6-AS1 ⁺ DP. Top: Dot plot of stage-specific DEGs (two-sided Wilcoxon rank-sum test, Bonferroni correction). Bottom left: MA plot of DEGs between 17 and 15 PCW (two-sided Wilcoxon rank-sum test, Bonferroni correction). Bottom right: GO enrichment analysis (one-sided hypergeometric test, Benjamini-Hochberg correction). h Chord diagram showing Pearson correlations among DLX6-AS1 , WNT/NOTCH pathway components, and odontogenic factors. Data in ( a – c , e – h ) are based on scRNA-seq and ST analyses from n = 2 biological replicates per developmental stage.

Article Snippet: Primary antibodies were applied against: DSPP (Abcam, ab216892), Ki67 (Cell Signaling Technology, 9129, Clone D3B5), KRT14 (Abcam, ab119695, Clone LL002), WNT6 (Abcam, ab50030), WNT7B (Proteintech, 10605-1-AP), WNT10B (Abcam, ab70816), LRP6 (Santa Cruz, sc-25317, Clone C-10), JAG1 (Abcam, ab300561, Clone EPR26388-51), NOTCH2 (Abcam, ab313453, Clone EPR28701-38), DKK1 (Abcam, ab61034, Clone EPR4759), SFRP1 (Proteintech, 26460-1-AP), HEY1 (Proteintech, 19929-1-AP), LEF1 (Abcam, ab137872, Clone EPR2029Y), BMP2 (Abcam,ab214821), BMP4 (Abcam, ab39973), GFP(CST, 2956, Clone D5.1) and rabbit IgG isotype control (CST, 3900, Clone DA1E) were added.

Techniques: Plasmid Preparation, Expressing, RNAscope, Staining

Journal: Nature Communications

Article Title: Spatiotemporal interplay between epithelial and mesenchymal cells drives human dentinogenesis

doi: 10.1038/s41467-026-69545-3

Figure Lengend Snippet: a Western blot analysis of DSPP, LEF1, and HEY1 in DLX6-AS1 ⁺ DMSC compared to the empty vector control (DMSC). b Mineralization assessment via ALP activity and Alizarin Red staining. c Analysis of the protein expression of LEF1 and HEY1 in DLX6-AS1 ⁺ DMSCs after treatment with WNTs, JAG1, or sequential WNTs-JAG1. Ctrl: DLX6-AS1 ⁺ DMSCs that received no treatment at all (i.e., no WNTs, JAG1, or standard mineralization induction). d Quantification of ALP and ARS staining in DLX6-AS1 ⁺ DMSCs following the stimulation protocols shown in ( c ). e IF analysis of DSPP (red) and KI67 (red) in DLX6-AS1 ⁺ DMSCs following the stimulation protocols shown in ( c ). Nuclei are counterstained with DAPI (blue). Scale bars: 100 μm. f Impact of epithelial WLS or JAG1 knockdown on DLX6-AS1 ⁺ DMSCs differentiation in co-culture system: protein levels of LEF1 and HEY1 by Western blot; DMP1 and DSPP expression by qPCR. g Western blot analysis of DSPP, LEF1, and HEY1 in DLX6-AS1 ⁺ DPSC compared to the empty vector control (DPSC). h Mineralization evaluation through ALP activity and Alizarin Red staining. i Analysis of the protein expression of LEF1 and HEY1 in DLX6-AS1 ⁺ DPSCs after treatment with WNTs, JAG1, or sequential WNTs-JAG1. Ctrl: DLX6-AS1 ⁺ DPSCs that received no treatment at all (i.e., no WNTs, JAG1, or standard mineralization induction). j IF analysis of DSPP (red) and KI67 (red) in DLX6-AS1 ⁺ DPSCs following the stimulation protocols in ( i ). Nuclei are counterstained with DAPI (blue). Scale bars: 100 μm. All experiments were independently repeated three times with similar results; representative images are shown for blots and staining. Data in all bar graphs are presented as mean ± SEM ( n = 3 independent experiments). Statistical significance for two-group comparisons ( a , b , g , h ) was determined by a two-sided unpaired Student’s t test. For multiple-group comparisons ( d , e , f , j ), a one-way ANOVA followed by Tukey’s multiple comparisons test was used. Exact P values are indicated on the graphs. A P value < 0.05 was considered statistically significant. Source data and full statistical details are provided in the Source Data file.

Article Snippet: Primary antibodies were applied against: DSPP (Abcam, ab216892), Ki67 (Cell Signaling Technology, 9129, Clone D3B5), KRT14 (Abcam, ab119695, Clone LL002), WNT6 (Abcam, ab50030), WNT7B (Proteintech, 10605-1-AP), WNT10B (Abcam, ab70816), LRP6 (Santa Cruz, sc-25317, Clone C-10), JAG1 (Abcam, ab300561, Clone EPR26388-51), NOTCH2 (Abcam, ab313453, Clone EPR28701-38), DKK1 (Abcam, ab61034, Clone EPR4759), SFRP1 (Proteintech, 26460-1-AP), HEY1 (Proteintech, 19929-1-AP), LEF1 (Abcam, ab137872, Clone EPR2029Y), BMP2 (Abcam,ab214821), BMP4 (Abcam, ab39973), GFP(CST, 2956, Clone D5.1) and rabbit IgG isotype control (CST, 3900, Clone DA1E) were added.

Techniques: Western Blot, Plasmid Preparation, Control, Activity Assay, Staining, Expressing, Knockdown, Co-Culture Assay