Journal: Biochemical Journal

Article Title: Heparan sulfate 6-O-endosulfatases: discrete in vivo activities and functional co-operativity

doi: 10.1042/bj20060848

Figure Lengend Snippet: Figure 6 Loss of mSulf activity increases the mitogenic response of MEFs to FGF2

Article Snippet: Recombinant human FGF2 and HGF/SF (HGF/scatter factor) were from R&D Systems (Abingdon, Oxon., U.K.).

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Multiple FGFR1 mutations modulate tumorigenic mechanisms in glioneuronal tumors

doi: 10.1101/2025.05.27.654799

Figure Lengend Snippet: R661P reduces K656E and N546K mutant protein stability by rescuing lysosomal degradation. A) Receptor stability experimental design: HEK293 cells expressing WT and mutant FGFR1 are stimulated with human FGF2 (hFGF2, 25 ng/mL) to induce receptor internalization and degradation and, in parallel, with cycloheximide (100 µg/mL) to block protein synthesis. B and D) Western blot analysis of a representative time-course experiment comparing WT FGFR1, K656E (B) and N546K (D) single and double mutants showing total Flag-FGFR1 protein levels at 0, 3 and 6 hours post-treatment with FGF2 and cycloheximide (CHX). C and E) Relative amounts of WT, single and double mutant FGFR1 protein obtained by quantifying western blots from n = 3 independent experiments. Values have been normalized (FC) against each relative 0h reference values. Data is represented by mean ± SEM and significant variations in protein amounts against each specific reference values have been indicated (ANOVA test, p value * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001) in the bar plots.

Article Snippet: Next, cells were induced with 25 ng/mL of recombinant human FGF2 (RD Systems) and treated with 100 µg/mL of Cycloheximide (Merck) and 200 nM of Bafilomycin A1 (Merck).

Techniques: Mutagenesis, Expressing, Blocking Assay, Western Blot

Journal:

Article Title: Serosal Mesothelium Retains Vasculogenic Potential

doi: 10.1002/dvdy.21334

Figure Lengend Snippet: Growth factors induce epithelial–mesenchymal transition (EMT) and smooth muscle actin (SMA) expression in serosal mesothelial cultures. Explants were cultured for 5 days under various conditions and analyzed for expression of Wt1 and SMA. A–C: After culture in normal medium, many peripheral cells of explants are fibroblastic and express SMA but not Wt1 (arrowheads), while Wt1-positive cells are negative for SMA (yellow open arrowhead). D–F: Under serum-free (sf) conditions, few of the epithelial sheet cells of cultured explants express SMA, whereas its majority express Wt1. SMA-positive cells can be Wt1-positive (open arrowheads) and Wt1-negative (arrowheads). G–L: Similar results were obtained for explants grown in sf-medium supplemented with basic fibroblast growth factor (bFGF) or epithelial growth factor (EGF), with few SMA-positive cells in the epithelial sheets. M–O: In explants grown in sf-medium supplemented with platelet-derived growth factor-BB (PDGF-BB), many fibroblastic peripheral cells in the epithelial sheet express SMA (arrowheads), many of which coexpress Wt1 (open arrowheads). Scale bar = 200 μm.

Article Snippet: Growth Factors Growth factors used were recombinant human EGF (40 ng/ml; R&D systems, 236-EG), recombinant human bFGF (40 ng/ml; R&D systems, 234-FSE/CF), and recombinant human homodimer PDGF-BB (40 ng/ml; Chemicon, GF018).

Techniques: Expressing, Cell Culture, Derivative Assay

Journal:

Article Title: Serosal Mesothelium Retains Vasculogenic Potential

doi: 10.1002/dvdy.21334

Figure Lengend Snippet: Production of smooth muscle actin (SMA) -positive cells in cultures of serosal mesothelium in the presence of specific growth factors. Cultures were analyzed at 5 days of culture with anti-SMA and anti-Wt1 antibodies. The total number of SMA-positive cells at the periphery was determined for each group indicated. One-way analysis of variance analysis determined that the number of SMA-positive cells in control serum-containing cultures was not significantly different from cultures with serum-free (sf) medium supplemented with platelet-derived growth factor-BB (PDGF-BB). In contrast to differentiation in serum-containing conditions, the number of SMA-positive cells in sf-medium or sf-medium supplemented with epithelial growth factor (EGF) or basic fibroblast growth factor (bFGF) was significantly (P < 0.001) reduced.

Article Snippet: Growth Factors Growth factors used were recombinant human EGF (40 ng/ml; R&D systems, 236-EG), recombinant human bFGF (40 ng/ml; R&D systems, 234-FSE/CF), and recombinant human homodimer PDGF-BB (40 ng/ml; Chemicon, GF018).

Techniques: Control, Derivative Assay