Journal: Molecular Cancer Research

Article Title: Demethylation of the SFRP4 Promoter Drives Gastric Cancer Progression via the Wnt Pathway

doi: 10.1158/1541-7786.mcr-20-0933

Figure Lengend Snippet: Figure 5. SFRP4 plays an antagonistic role on biology function of SFRP1. A, The expression of Wnt ligands (Wnt2, Wnt3a, Wnt4, Wnt5A) in different gas- tric cancer cells was examined by Western blotting. B, The binding activ- ity of Wnt5A to its receptor was exam- ined by flow cytometry in the absence or presence of recombinant SFRP4. C, Co-IP experiments in MGC80–3cells were performed using anti-SFRP4 or anti-b-catenin antibodies in the absence or presence of recombinant Wnt5a. IgG was used as a negative control. D, The Wnt pathway markers (GSK-3b, b-catenin, c-Jun, LEF1, CD44, and MMP7) were examined in MGC80–3 cell after treating with recombinant SFRP1 (100 ng/mL), recombinant SFRP4 (100 ng/mL), or both for 48 hours. E, Afterward, com- bination of recombinant SFRP1 and recombinant SFRP4 were addedto the MGC80–3 cell in a concentration gra- dient (1:0, 1:0.5, 1:1, 1:2, 1:3) for 48 hours before harvesting for Western blotting analysis. F, The effects of recombinant SFRP1 and SFRP4 on the proliferation of MGC80–3 and AGS cellswere exam- ined by CCK-8 assay. , P < 0.001. r, recombinant; NC, negative control. In (A–E), the images were representa- tive of three independent experi- ments. In F, the experiments were repeated three times. Statistical sig- nificance was determined by one-way ANOVA test (F).

Article Snippet: The recombinant human SFRP1 (1384-SF), SFRP4 (1827-SF) and Wnt5a(645-WN) protein were purchased from R&D Systems.

Techniques: Expressing, Western Blot, Binding Assay, Cytometry, Recombinant, Co-Immunoprecipitation Assay, Negative Control, Concentration Assay, CCK-8 Assay

Journal: Scientific Reports

Article Title: Secreted frizzled-related protein 1 overexpression in gastric cancer: Relationship with radiological findings of dual-energy spectral CT and PET-CT

doi: 10.1038/srep42020

Figure Lengend Snippet: ( a ) In vitro angiogenesis assays were performed by treating HUVEC cells with culture supernatants of SGC-7901/vector or SGC-7901/sFRP1 cells. Representative pictures are shown (original magnification,x40). Numbers of tube formation per field are plotted. The data are shown as mean ± SD of three independent experiments ( p < 0.05). ( b ) Migration of HUVECs was conducted using Boydon chamber assays. Same amount of SGC-7901/vector and SGC-7901/sFRP1 cells were plated in 24-well plate for 24 h, and the culture supernatants were collected and used as chemoattractants to HUVECs in the migration assays. Representative pictures are shown (left, original magnification,x100) and migrated cell numbers are plotted (right). The data are shown as mean ± SD of three independent experiments ( p < 0.05).

Article Snippet: Plasmid containing Myc-DDK-tagged ORF clone of Homo sFRP1 was purchased from Origene (Origene Technologies, Rockville, MD).

Techniques: In Vitro, Plasmid Preparation, Migration

Journal: Scientific Reports

Article Title: Secreted frizzled-related protein 1 overexpression in gastric cancer: Relationship with radiological findings of dual-energy spectral CT and PET-CT

doi: 10.1038/srep42020

Figure Lengend Snippet: ( a ) In vitro angiogenesis assays were performed by treating HUVEC cells with culture supernatants of SGC-7901/vector or SGC-7901/sFRP1 cells. Representative pictures are shown (original magnification,x40). Number of tube formation per field are plotted. ( b ) Migration of HUVECs was conducted using Boydon chamber assays. Same amount of SGC-7901/vector and SGC-7901/sFRP1 cells were plated in 24-well plate for 24 h, and the culture supernatants were collected and used as chemoattractants to HUVECs in the migration assays. Representative pictures are shown (left, original magnification,x100) and migrated cell numbers are plotted (right). ( c ) VEGF levels in culture supernatants supernatants of SGC-7901/vector and SGC-7901/sFRP1 cells were examined using ELISA analysis.

Article Snippet: Plasmid containing Myc-DDK-tagged ORF clone of Homo sFRP1 was purchased from Origene (Origene Technologies, Rockville, MD).

Techniques: In Vitro, Plasmid Preparation, Migration, Enzyme-linked Immunosorbent Assay

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Wnt1 Boosts Fracture Healing by Enhancing Bone Formation in the Fracture Callus.

doi: 10.1002/jbmr.4797

Figure Lengend Snippet: Fig. 7. Effects of recombinant Wnt1 on critical-size bone defect healing. (A) Experimental design. (B) Representative 3D reconstructions from μCT datasets. The entire area between the two inner pin holes in shown. Red indicates less mineralized bone, whereas yellow and blue indicate higher mineralized bone. (C) Bone volume in the defect area. (D) Polar moment of inertia of the bone in the defect area as analyzed by μCT evaluation. (E) Trabecular thickness, (F) trabecular separation, and (G) trabecular number of the newly formed bone in the defect area. (H) Active YAP1 and BMP2 immunostaining in the defect area. Scale bar = 100 μm. n = 7 per group. Mann-Whitney U test. sFRP1, secreted frizzled-related protein 1.

Article Snippet: The critical-size model has been described. (31) Five micrograms (5 μg) of recombinant sFRP1 protein alone (R&D Systems; 9019-SF-025) was used for the control group.

Techniques: Recombinant, Immunostaining, MANN-WHITNEY