recombinant human wnt10b  (R&D Systems)

Journal: Stem Cell Research & Therapy

Article Title: Synergistic effects of mesenchymal stem cell secretome and Wnt10b on hair follicle regeneration in a 3D bioengineering model: a preclinical study

doi: 10.1186/s13287-025-04830-4

Figure Lengend Snippet: Hair follicle-specific markers expression. A) The representative illustration of the immunofluorescence staining of DPC (ALP, COL1, α-SMA, Versican) and epithelial markers (K75, K15, AE13, AE15, K71) in control group and the Wnt10b + secretome group. B) Semi-quantitative measurement of protein expression on the generated follicles. C) Expression levels of hair growth-related genes after treatment by PMSCs-derived secretome and Wnt10b compared to control group (as fold change). Data are shown as mean + rang. Data are gathered from at least 5 random follicles from each samples for total of 3 samples per group

Article Snippet: The medium was composed of: (A) changing media alone (control group), (B) changing media with 1 mg/mL secretome, (C) changing media with 1 μg/mL recombinant human Wnt10b (R&D Systems), or (D) changing media with secretome and Wnt10b.

Techniques: Expressing, Immunofluorescence, Staining, Control, Generated, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Synergistic effects of mesenchymal stem cell secretome and Wnt10b on hair follicle regeneration in a 3D bioengineering model: a preclinical study

doi: 10.1186/s13287-025-04830-4

Figure Lengend Snippet: Transplantation of hydrogel containing microfollicles into skin of nude mice. A) GelMA hydrogels containing hair microfollicles from Wnt10b + secretome group or control group were transplanted under the dorsal skin of nude mice. Nude mice are shown before and after transplantation of hydrogel containing microfollicles. B-I) Two weeks post-transplantation, the hydrogels were surgically excised, and the transplantation sites were evaluated through transverse and longitudinal histological analyses. B-C) H&E staining of transverse and longitudinal sections of skin samples. D) Comparison of hair follicle number between control group and Wnt10b + secretome group. E) Comparison of follicle length between control group and Wnt10b + secretome group. F-G) Masson’s Trichrome staining of both transverse and longitudinal sections of skin samples to evaluate the collagen deposition. H) Comparison of collagen deposition in transverse sections between control group and Wnt10b + secretome group. I) Comparison of collagen deposition in longitudinal sections between control group and Wnt10b + secretome group Data are illustrated as Mean ± SD. Data are gathered from at least 5 random follicles from each samples for total of 3 samples per group

Article Snippet: The medium was composed of: (A) changing media alone (control group), (B) changing media with 1 mg/mL secretome, (C) changing media with 1 μg/mL recombinant human Wnt10b (R&D Systems), or (D) changing media with secretome and Wnt10b.

Techniques: Transplantation Assay, Control, Staining, Comparison

Journal: Stem Cell Research & Therapy

Article Title: Synergistic effects of mesenchymal stem cell secretome and Wnt10b on hair follicle regeneration in a 3D bioengineering model: a preclinical study

doi: 10.1186/s13287-025-04830-4

Figure Lengend Snippet: Immunofluorescence staining and RT-PCR analysis of skin samples after transplantation of hydrogel containing microfollicles. (A) GelMA hydrogels containing DPCs and epithelial cells (pre-treated with Wnt10b + secretome or control) were transplanted subcutaneously into nude mice. Two weeks post-transplantation, the hydrogels were surgically excised, and the graft sites were analyzed via immunofluorescence staining. The representative illustration of the immunofluorescence staining of DPC (ALP, COL1, α-SMA, Versican) and epithelial markers (K75, K15, AE13, AE15, K71) in control group and the Wnt10b + secretome group. (B) Semi-quantitative measurement of protein expression on the generated follicles. (C) Expression levels of hair growth-related genes after treatment by PMSCs-derived secretome and Wnt10b compared to control group (as fold change). Data are shown as mean + rang. Data are gathered from at least 5 random follicles from each samples for total of 3 samples per group

Article Snippet: The medium was composed of: (A) changing media alone (control group), (B) changing media with 1 mg/mL secretome, (C) changing media with 1 μg/mL recombinant human Wnt10b (R&D Systems), or (D) changing media with secretome and Wnt10b.

Techniques: Immunofluorescence, Staining, Reverse Transcription Polymerase Chain Reaction, Transplantation Assay, Control, Expressing, Generated, Derivative Assay

Journal: Cell death & disease

Article Title: Alternative Wnt-signaling axis leads to a break of oncogene-induced senescence.

doi: 10.1038/s41419-024-06550-8

Figure Lengend Snippet: Fig. 4 Role of Wnt10b on OIS and in melanoma cells. A Representative immunohistochemical staining of Wnt10b protein in normal human skin, primary human melanoma, and human metastatic melanoma tissue samples (n = 10). B Percentages of SA-β-Galactosidase positive cells (blue) in mock/BRAFm-transduced NHEMs treated with recombinant Wnt10b or PBS (left) (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). Example image of light microscopic examination of SA-β-Galactosidase staining (right). C Immunofluorescence staining’s of PML and DAPI in mock/BRAFm-transduced NHEMs treated with recombinant Wnt10b or PBS. The graph shows nuclear accumulation of PML (left) (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). Sample image of overlays of PML (red) and DAPI (blue) (right). D Representative real-time cell proliferation curves of BRAFm-transduced NHEMs treated with recombinant Wnt10b or PBS (BRAFm PBS set as 1) (right panel) and quantified “slope” (proliferative ability) (left panel) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). E Percentages of SA-β-Galactosidase positive cells (blue) in SBcl2, WM1366, and MV3 cell lines 48 h after transfection with siWnt10b or siCtr (left) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). Example image of light microscopic examination of SA- β-Galactosidase staining in SBcl2 cells (right). F Immunofluorescence staining of PML and DAPI in melanoma cell lines 48 h after the transfection with siWnt10b or siCtr. The graph shows the number of nuclear PML bodies (left) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). Sample image of overlays of PML (red) and DAPI (blue) in SBcl2 cells (right). G Cell cycle analysis 48 h after Wnt10b inhibition in SBcl2, WM1366, and MV3 cells. Bars represent cells in G1 phase (left) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). Representative histograms of cell cycle analysis in SBcl2 cells 48 h after siWnt10b or siCtr transfection.

Article Snippet: Seven days after viral transfection, NHEMs were treated with recombinant Wnt10b (200 ng/ml; R&D Systems, Inc., Minneapolis USA) with PBS as control and AMBMP hydrochloride (50 nM; R&D Systems) with DMSO as control.

Techniques: Immunohistochemical staining, Staining, Recombinant, Transfection, Cell Cycle Assay, Inhibition

Journal: International Journal of Molecular Sciences

Article Title: Immunocytochemical Analysis of Endogenous Frizzled-(Co-)Receptor Interactions and Rapid Wnt Pathway Activation in Mammalian Cells

doi: 10.3390/ijms222112057

Figure Lengend Snippet: Wnt10B-dependent activation of the canonical Wnt/β-catenin pathway following FZD6–LRP6 co-localization. Co-localization of FZD6 and LRP6 by Wnt10B treatment and subsequent detection of nuclear β-catenin indicates rapid receptor interaction and early Wnt/β-catenin signaling in PC-3. ( a – f ) Detection of FZD6–LRP6 interaction complexes by PLA after 2 and 4 min of Wnt10B incubation ( b , e ) versus vehicle-treated controls ( a , d ). FZD6–LRP6 interaction complexes were frequently observed in the nuclei (circles). FZD6–LRP6 complexes (red); plasma membranes (green); nuclei (blue). Scale bar: 10 µm. ( c , f ) Quantification of PLA signal density by particle analyses at the single-cell level. Particles were normalized to the area of cells (ROIs). The amount of FZD6–LRP6 complexes was significantly higher after both 2 min ( n = 317 vs. n = 297 cells) and 4 min ( n = 264 vs. n = 263 cells, vehicle vs. Wnt10B) of Wnt10B incubation (* p ≤ 0.05, Mann–Whitney Test); mean + SD; n = 3 independent experiments. ( g – k ) Well-documented translocation of β-catenin into the nucleus. ( g – i ) Immunofluorescence images of β-catenin staining. ( g ) Membrane association of β-catenin in control cells; ( h , i ) nuclear localization of β-catenin after incubation of PC-3 cells for 20 min with Wnt10B; arrows indicate nuclear β-catenin labeling. β-Catenin (red); nuclei (blue). Scale bar: 10 µm. ( j , k ) Quantification of β-catenin (FI, fluorescence intensity) in DAPI-positive nuclei. Significantly increased nuclear β-catenin was detected within 2 to 5 min of Wnt10B incubation ( j ) and continued until 60 min of incubation ( k ); * significance vs. vehicle-treated control (* p ≤ 0.05, Mann–Whitney Test); ( k ) nuclear β-catenin (normalized to vehicle-treated control) followed a linear trend until 40 min of Wnt10B incubation (# p ≤ 0.05, one-way ANOVA, post hoc test for linear trend, R 2 = 0.915); mean + SD; n = 3 independent experiments.

Article Snippet: Recombinant Wnt5A and Wnt10B peptides were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, Incubation, Clinical Proteomics, MANN-WHITNEY, Translocation Assay, Immunofluorescence, Staining, Membrane, Control, Labeling, Fluorescence