Fig 1: SCRG1 regulated chondrogenesis of UCMSCs through Wnt5a signaling pathway. (a) Regulating pluripotency of stem cell signaling pathway was the most significantly enriched KEGG pathway based on GSEA, wnt5a was negatively correlated with this pathway after SCRG1 overexpression. (b, c) Western blot analysis showed that Wnt5a decreased after shSCRG1 transfection and was restored after administration of rhSCRG1 (450 ng/mL); protein levels of β-catenin exhibited the opposite trend to Wnt5a. (d, e) Protein levels of COL2A1 and ACAN were detected in UCMSCs by IHC. The results showed that the protein levels of COL2A1 and ACAN were reduced following SCRG1 knockdown and restored after treated with rhWnt5a (300 ng/mL) and rhSCRG1 (450 ng/mL) protein at 24 h. Scale bar = 100 μm. Experiments were repeated three times, ∗P < 0.05 (t-test).
Fig 2: Heterogeneous Fibroblast Populations in Injured and Uninjured Neonatal Hearts(A) UMAP representation of different FB sub-populations analyzed.(B) Heatmap showing expression of top enriched genes for each FB sub-population.(C) Percentage of each FB sub-population over total nonmyocytes within each sample.(D–H) UMAP plots showing expression of Pro.FB-enriched gene Hmgb2 (D), FBI-enriched gene Cxcl1 (E), FB2-enriched gene Dlk1 (F), FB3-enriched gene Nov (G), and FB4-enriched gene Fbln5 (H).(I) Heatmap showing relative fold induction (Z score) of Serpinb2, Wnt5a, and Ltbp3 expression at various time points after P1 or P8 MI detected by bulk RNA-seq.(J) EdU incorporation (magenta) and vimentin immunofluorescent staining (green) of NRCF cells treated with 200 ng/mL BSA (negative control), 20 ng/mL recombinant SERPINB2, 100 ng/mL recombinant LTBP3, or 100 ng/mL recombinant WNT5A (positive control), with quantification showing the proportion of EdU-positive cells among vimentin-positive cells (fibroblasts) (n = 4 per each group; ****p < 0.0001). Scale bar, 100 µm.See also Figure S6.
Fig 3: Wnt3a induces odontoblastic differentiation of DPSCs. a Uniform manifold approximation and projection (UMAP) of single-cell transcriptomic sequencing data from DPSCs treated with blank, Wnt3a, Wnt5a, and Wnt10a under odontogenic induction conditions. b The correlation heatmap of scRNA-seq datasets. c Volcano plot illustrating the differentially expressed genes (DEGs) of Wnt3a, Wnt5a, and Wnt10a-treated groups compared with control group. d Gene Ontology (GO) enrichment analysis of upregulated DEGs in Wnt3a-treated groups. Alkaline phosphatase (ALP) staining at day 7 (e) and Alizarin Red S (ARS) staining at day 21 (f) post-treatment. g Protein expression and quantitative analysis of odontoblast differentiation-related markers in DPSCs after 7-day treatment. (Data are presented as the mean of >3 biological replicates ± SD. ****P < 0.000 1)
Fig 4: Wnt3a induces formation of an NKD1+ subpopulation with odontoblastic differentiation potential in DPSCs. a Cluster-resolved UMAP of scRNA-seq data from DPSCs treated with blank, Wnt3a, Wnt5a, and Wnt10a under odontogenic conditions. b Bar plot showing the proportional distribution of identified clusters. c GO enrichment analysis of upregulated genes of cluster 3. d FeaturePlot visualization of NKD1 expression pattern. e Immunofluorescence analysis of NKD1 co-localization with DSPP. White arrowheads indicate cells positive for both NKD1 and DSPP. f Flow cytometry quantification of NKD1+DSPP+ cell proportions. (Data are presented as the mean of >3 biological replicates ± SD. ****P < 0.000 1)
Fig 5: CELSR2 KD compromises WNT3A-induced proliferation of U87 MG cells.A, B Administration of WNT3A, WNT5A and WNT1 significantly enhanced cell proliferation of cultured glioma cells, as revealed by EDU staining. C, D In CELSR2-KD U87 MG cells, WNT5A and WNT1, but not WNT3A, significantly enhanced cell proliferation as identified by EDU labeling. E, F Flow cytometry showed a decrease in the G0/G1 ratio and an increase in the S phase in the control+WNT3A group (U87 MG cells treated with WNT3A) compared to the control group (U87 MG cells), and no significant change in the KD group (CELSR2-KD U87 MG cells) compared to the KD + WNT3A group (CELSR2-KD U87 MG cells treated with WNT3A). G, H Western blots showed a significant increase of β-catenin and cyclin D1, and a significant decrease in GSK-3β in the Ctrl+WNT3A group compared to the control group, and there was no significant protein change in the KD group compared to the KD + WNT3A group. Scale bar is 50 μm in (A, C). Data are represented as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; n = 3, Student’s t-test.
Supplier Page from Abcam for Recombinant Human Wnt5a protein