sfrp1  (R&D Systems)

Journal: Nature Communications

Article Title: The pericardium forms as a distinct structure during heart formation

doi: 10.1038/s41467-025-63599-5

Figure Lengend Snippet: A – E Ventral confocal imaging (max projection) of representative 72 hpf hand2:EGFP;myl7DsRed larvae undergoing distinct treatments as indicated; anterior to the top; v ventricle, a atrium. A , B Representative larvae treated with DMSO vehicle only as control at 18 hpf overnight showing hand2:EGFP- expressing pericardial sac surrounding the heart at 72 hpf ( A , 20x) and cellular density ( B , 40x zoom, representative nuclei marked with dashed lines). C , D Ventral images of representative hand2:EGFP;myl7:DsRed larvae treated with the Wnt signaling inhibitor IWR-1 at 18 hpf overnight, showing expanded pericardial sac and edema with large, stretched cells surrounding the larval zebrafish heart at 72 hpf ( C 20x) and lower cellular density ( D 40x). E , F Ventral images of hand2:EGFP;myl7:DsRed larvae treated with BDM as myosin II inhibitor at 18 hpf overnight showing expanded pericardial sac with normal cell distribution at 72 hpf ( E 20x) and cellular density ( F 40x). G – J Quantifications of pericardial and cardiac features following the treatments. One-way ANOVA, n = 6 animals, three independent experiments. G Heart rate of vehicle-treated, Wnt-inhibited, and myosin II-inhibited (BDM) animals ( p = 0.6056 DMSO to IWR-1, p = 0.0001 DMSO to BDM). H Pericardial area (distribution per ventral view), showing increased pericardial area in IWR-1-treated animals ( p = 0.0631 DMSO to IWR-1, p = 0.317- DMSO to BDM). I Cell density (cells per square millimeter), showing decreased cell density in IWR-1-treated animals only ( p = 0.0001 DMSO to IWR-1, p = 0.01345 DMSO to BDM). J Cell size showing increases in IWR-1-treated embryos only ( p = 0.0001 DMSO to IWR-1, p = 0.9918 DMSO to BDM). K , L Increased tissue stiffness in the pericardium of rats treated with PBS (vehicle), Iso only, sFRP1 only, or combined Isoproterenol (Iso) and SFRP1 ( n = 3 per condition). Neonatal rats (0-to-4-day old rats) were injected intraperitoneally with 0.05 mg/kg/day of human recombinant sFRP1 protein and Iso in an animal model of pediatric dilated cardiomyopathy. Atomic force microscopy (AFM) of dissected pericardia provided measures for Young’s modulus (kPa) as readout for tissue elasticity, with treated pericardia showing increased stiffness with combined Iso and sFRP1 only ( K ) as quantified per sample( L , unpaired two-tailed t -test, p = 0.9093 vehicle to Iso only, p = 0.6129 vehicle to sFRP1 only, p = 0.0140 vehicle to Iso + sFRP1). Each dot represents an individual sample. Representative images of control and = treated rats. Source data are provided as a Source Data file. Scale bar A , C , E 200 μm; B , D , F (40x) 50 μm. Species silhouettes were adapted from the PhyloPic database ( https://www.phylopic.org/ ).

Article Snippet: The sFRP1 (Recombinant Human sFRP1 Protein, CF, R&D systems) solution was freshly prepared for each treatment by dissolving in phosphate-buffered saline (PBS) at room temperature.

Techniques: Imaging, Control, Expressing, Injection, Recombinant, Animal Model, Microscopy, Two Tailed Test

Journal: Frontiers in Cell and Developmental Biology

Article Title: SFRP1 induces a stem cell phenotype in prostate cancer cells

doi: 10.3389/fcell.2023.1096923

Figure Lengend Snippet: (A) Stratification of PRAD samples. Samples were clustered based on the expression of FAD and ACTA2 genes. A 3-cluster configuration was obtained with partition around medoid (PAM) algorithm from the Consensus Clustering Analysis. Heatmap shows the level of expression of both genes across samples in clusters. (B) Profile of differentially expressed genes between clusters. The heatmap shows the expression of differentially expressed genes between clusters of samples defined. Significant differences in gene expression were assessed and determined with a Fold-change >2 and adjusted p -value <0.05. (C) Volcano plot for differentially expressed genes between clusters, highlighting SFRP1 and SOX2 overexpression. (D) Differences in cell type enrichment between clusters. Comparison between the enrichment scores distributions of selected cell types are shown in the box and whiskers plot. Significant differences were determined with a two-sided t -test. Asterisk indicates differences with a p -value <0.05. (E) Significantly enriched KEGG terms in differentially expressed genes between cluster_3 and cluster_1 (relevant pathways in cancer). (F) qPCR for SFRP1 transcript in normal fibroblasts (NFs), cancer-associated fibroblasts (CAFs), LNCaP, and DU145 cell lines (* p -value<0.05). (G) Western blot for SFRP1 protein in NFs, CAFs, LNCaP, and DU145 cell lines. (H) CM-CAFs; CM-LNCaP, and CM-Du145 represent SFRP1 in conditioned medium of each cell line.

Article Snippet: The treatments were performed with SFRP1 protein (R&D cat.5396-SF-025) at 0.1 nM with PBS carrier buffer as vehicle.

Techniques: Expressing, Gene Expression, Over Expression, Comparison, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: SFRP1 induces a stem cell phenotype in prostate cancer cells

doi: 10.3389/fcell.2023.1096923

Figure Lengend Snippet: (A) LNCaP and DU145 cells were incubated for 48 h with SFRP1 or vehicle. Expression levels of SOX2, OCT4, and NANOG were determined by qPCR. Data shows relative mRNA expression; ß -actin (LNCaP) and 18S (DU145) were used as housekeeping gene (* p < 0.05). (B) Protein levels were determined by Western blot, lamin b1 was used as loading control, (* p < 0.05).

Article Snippet: The treatments were performed with SFRP1 protein (R&D cat.5396-SF-025) at 0.1 nM with PBS carrier buffer as vehicle.

Techniques: Incubation, Expressing, Western Blot, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: SFRP1 induces a stem cell phenotype in prostate cancer cells

doi: 10.3389/fcell.2023.1096923

Figure Lengend Snippet: (A) Flow cytometry analysis of the CD44+/CD133+ subpopulation in cells treated with SFRP1 or vehicle. (B) Representative images of tumorspheres formed in LNCaP and DU145 cells treated with SFRP1 or vehicle; tumorspheres were counted from 1,000 seeding cells, and the volume was calculated with the equation V = 4/3 πr³ (* p < 0.05). (C) The χ2 tests were calculated using ELDA software to evaluate the frequency of PCSCs. The number of cells plated is displayed against the log proportion of wells devoid of tumorspheres. The tables illustrates the estimated frequency of PCSCs.

Article Snippet: The treatments were performed with SFRP1 protein (R&D cat.5396-SF-025) at 0.1 nM with PBS carrier buffer as vehicle.

Techniques: Flow Cytometry, Software

Journal: Frontiers in Cell and Developmental Biology

Article Title: SFRP1 induces a stem cell phenotype in prostate cancer cells

doi: 10.3389/fcell.2023.1096923

Figure Lengend Snippet: (A) Cell apoptosis of LNCaP and DU145 was detected by flow cytometry. Cells were treated with either SFRP1 or vehicle (CTRL); FITC-Annexin V coupled antibody and propidium iodide was used for the detection. (B) MTT assay was used to determine cytotoxicity. LNCaP and DU145 cells were treated for 72 h with bicalutamide (50 μM) or bicalutamide + SFRP1 or vehicle, (* p < 0.05). (C) Proliferation curve was perform using MTT assay kit. LNCaP and DU145 cells were treated for 24, 48, 72 and 96 h with: Bicalutamide (50 μM), bicalutamide + SFRP1 or vehicle, every 24 h the O. D (590 nm) was obtained (* p < 0.05).

Article Snippet: The treatments were performed with SFRP1 protein (R&D cat.5396-SF-025) at 0.1 nM with PBS carrier buffer as vehicle.

Techniques: Flow Cytometry, MTT Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: SFRP1 induces a stem cell phenotype in prostate cancer cells

doi: 10.3389/fcell.2023.1096923

Figure Lengend Snippet: (A) The clonogenic ability of LNCaP and DU145 was analyzed. Cells were treated with SFRP1 or vehicle for 2 weeks, after incubation, cells were stained with crystal violet and analyzed using ImageJ software. (B) Cell migration of LNCaP and DU145 cells was detected by a wound healing assay. Cells were treated with either SFRP1 or vehicle for 48 h before their evaluation (C) . Cell invasion of LNCaP and DU145 cells was detected by the Transwell assay. Cells were treated with either SFRP1 or vehicle and the invading cells were counted (* p 0.05).

Article Snippet: The treatments were performed with SFRP1 protein (R&D cat.5396-SF-025) at 0.1 nM with PBS carrier buffer as vehicle.

Techniques: Incubation, Staining, Software, Migration, Wound Healing Assay, Transwell Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: SFRP1 induces a stem cell phenotype in prostate cancer cells

doi: 10.3389/fcell.2023.1096923

Figure Lengend Snippet: LNCaP and DU145 cell lines were treated with SFRP1; SFRP1 + ICRT14 or vehicle for 48 h. (A) The differential expression of WNT target genes was evaluated using qRT–PCR. Actin-b served as the normalization control. (B) Western blot was used to measure the protein levels of AXIN2 and MYC. The relative expression was quantified using ImageJ software and is displayed below the western blots (fold). (C) Luciferase reporter assay of ß -catenin-Tcf/Lef activity was used. LNCaP and DU145 cells were transfected with the Wnt signalling reporter (TOP flash or the control FOP flash). It shows the TOP/FOP fold change relative to control (* p < 0.05).

Article Snippet: The treatments were performed with SFRP1 protein (R&D cat.5396-SF-025) at 0.1 nM with PBS carrier buffer as vehicle.

Techniques: Quantitative Proteomics, Quantitative RT-PCR, Control, Western Blot, Expressing, Software, Luciferase, Reporter Assay, Activity Assay, Transfection

Journal: Frontiers in Cell and Developmental Biology

Article Title: SFRP1 induces a stem cell phenotype in prostate cancer cells

doi: 10.3389/fcell.2023.1096923

Figure Lengend Snippet: The suggested mechanism by which the loss of SFRP1 expression in epithelial cells linked with the stromal SFRP1 signalling axis promotes the stemness of prostate cancer cells and reduces their sensitivity to bicalutamide. SFRP1 induces higher SOX2, OCT4, and NANOG expression levels, all within the context of WNT/β-catenin pathway activation.

Article Snippet: The treatments were performed with SFRP1 protein (R&D cat.5396-SF-025) at 0.1 nM with PBS carrier buffer as vehicle.

Techniques: Expressing, Activation Assay