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: International journal of molecular medicine

Article Title: Enhanced rapamycin delivery to hemangiomas by lipid polymer nanoparticles coupled with anti-VEGFR antibody.

doi: 10.3892/ijmm.2018.3518

Figure Lengend Snippet: Figure 7. VEGF‑A and bFGF production in (A and B) HUVECs and (C and D) HemECs analyzed by ELISA. The relative protein levels in each treatment group were expressed as a % relative to the untreated group. Data are expressed as mean ± standard deviation (n=3). *P<0.05, **P<0.01 and ***P<0.001, with compari- sons indicated by brackets. VEGF, vascular endothelial growth factor; bFGF, basic fibroblast growth factor; HUVECs, human umbilical vein endothelial cells; HemECs, hemangioma endothelial cells; PLNPs‑V, lipid polymer nanoparticles coupled with anti‑VEGR2 antibody; R‑PLNPs, rapamycin‑encapsulated lipid polymer nanoparticles; R‑PLNPs‑V, R‑PLNPs coupled with anti‑VEGR2 antibody.

Article Snippet: The mouse anti-human VEGFR2 monoclonal antibody (MAB3571; 1:1,000), and the basic fibroblast growth factor (bFGF) (dFB50) and VEGF-A (dVE00) ELISA kits were purchased from R&d Systems, Inc. (Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Polymer

Journal: Cell stem cell

Article Title: A Non-Coding Disease Modifier of Pancreatic Agenesis Identified by Genetic Correction in a Patient-Derived iPSC Line

doi: 10.1016/j.stem.2020.05.001

Figure Lengend Snippet: Key Resources Table:

Article Snippet: Recombinant human bFGF , R & D Systems , 233-FB/CF.

Techniques: Recombinant, Knock-Out, Reporter Assay, Plasmid Preparation, Cloning, Quantitative RT-PCR, Software

Journal: Brain : a journal of neurology

Article Title: Improvement in disability after alemtuzumab treatment of multiple sclerosis is associated with neuroprotective autoimmunity.

doi: 10.1093/brain/awq176

Figure Lengend Snippet: Figure 3 Neurotrophin secretion from PBMC cultures from 15 patients before and 12 months after alemtuzumab treatment. Cultures were either unstimulated (unstim) or stimulated with the myelin antigen, MBP or polyclonally stimulated with anti-CD3/anti-CD28 antibodies (CD3/28). Supernatants were harvested after 72 h and assayed for: (A) brain-derived neurotrophic factor, (B) CNTF, (C) platelet-derived neurotrophic factor, (D) FGF, and (E) insulin-like growth factor-1. Error bars represent 95% confidence intervals. (*P50.05, **P50.01, ***P50.001).

Article Snippet: For blocking experiments, neutralizing antibodies to BDNF (2 mg/ml—Sigma B5050), CNTF (0.2 mg/ml—Peprotech 500-P140), FGF (2 mg/ml RnD systems AB-233-NA) and PDGF (20 mg/ml RnD systems AN-20-NA) were added at the time of resuspension in PBMC derived conditioning medium.

Techniques: Derivative Assay

Journal: Brain : a journal of neurology

Article Title: Improvement in disability after alemtuzumab treatment of multiple sclerosis is associated with neuroprotective autoimmunity.

doi: 10.1093/brain/awq176

Figure Lengend Snippet: Figure 4 Peripheral blood mononuclear cell neurotrophin secretion from three patients 12 months after alemtuzumab in response to a wide variety of antigens. PBMCs were cultured either unstimulated (unstim) or stimulated with; MBP, tetanus toxoid (TT), myelin oligodendrocyte glycoprotein (MOG), collagen type II fragment aa245–270 (Collagen), myelin basic peptide aa87–99 (MBP-P), keyhole limpet haemocyanin (KLH) or recombinant human insulin (Insulin). Supernatants were harvested after 72 h and assayed for: (A) brain-derived neurotrophic factor, (B) CNTF, (C) platelet-derived neurotrophic factor, (D) insulin-like growth factor-1 and (E) FGF (*P50.05).

Article Snippet: For blocking experiments, neutralizing antibodies to BDNF (2 mg/ml—Sigma B5050), CNTF (0.2 mg/ml—Peprotech 500-P140), FGF (2 mg/ml RnD systems AB-233-NA) and PDGF (20 mg/ml RnD systems AN-20-NA) were added at the time of resuspension in PBMC derived conditioning medium.

Techniques: Cell Culture, Recombinant, Derivative Assay

Journal: Brain : a journal of neurology

Article Title: Improvement in disability after alemtuzumab treatment of multiple sclerosis is associated with neuroprotective autoimmunity.

doi: 10.1093/brain/awq176

Figure Lengend Snippet: Figure 5 Peripheral blood mononuclear cell neurotrophin secretion, induced by MBP stimulation. PBMCs from 15 healthy controls (HC), 15 patients before (pre) and at three time points (6, 9 and 12 months) after alemtuzumab and from 10 patients treated with interferon b-1a (rebif), were cultured with MBP. Supernatants were harvested after 72 h and assayed for: (A) brain-derived neurotrophic factor, (B) CNTF, (C) platelet-derived neurotrophic factor, (D) insulin-like growth factor-1 and (E) FGF. Error bars represent 95% confidence intervals (**P50.01, ***P50.001).

Article Snippet: For blocking experiments, neutralizing antibodies to BDNF (2 mg/ml—Sigma B5050), CNTF (0.2 mg/ml—Peprotech 500-P140), FGF (2 mg/ml RnD systems AB-233-NA) and PDGF (20 mg/ml RnD systems AN-20-NA) were added at the time of resuspension in PBMC derived conditioning medium.

Techniques: Cell Culture, Derivative Assay

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

Journal: Stem cell research & therapy

Article Title: Differentiation of adipose-derived stem cells into Schwann cell-like cells through intermittent induction: potential advantage of cellular transient memory function.

doi: 10.1186/s13287-018-0884-3

Figure Lengend Snippet: Fig. 1 Identification of adipose-derived stem cells (ASCs) and schematic of the experimental design. a Primary ASCs grew in clusters and had a rounded spindle-like shape. b ASCs at passage 2 could differentiate into adipocytes, and the visual field in photographs was filled with red lipid droplets stained with Oil Red O solution. c ASCs differentiated into osteocytes formed calcium nodules with burrs, which were stained red by Alizarin Red solution. d In cartilage pellets, many cartilage lacunae were found, and the glycosaminoglycans around the chondrocytes were stained purple-blue by Toluidine Blue O solution. e Flow cytometry showed that more than 98% of ASCs were immunopositive for the MSC markers CD29, CD44, and CD90, while less than 6% of ASCs were immunopositive for the hematopoietic stem cell markers CD34, CD45, and CD86. f Schematic showing the experimental groups and induction methods. bFGF, basic fibroblast growth factor; β-ME, β-mercaptoethanol; dASC, differentiated ASC; DMEM, Dulbecco’s modified Eagle’s medium; FBS, fetal bovine serum; PDGF, platelet-derived growth factor; SC, Schwann cell; uASC, undifferentiated ASC

Article Snippet: Pre-induction medium I was replaced with pre-induction medium II consisting of DMEM, 10% FBS, and 35 ng/ml all trans-retinoic acid (RA) (R2625; Sigma) and incubated for 72 h. The resulting differentiated ASCs were divided into four groups according to subsequent processing: (ii) sustaining dASCs 4d: Differentiated ASCs (dASCs) were induced for 4 days with complete induction medium consisting of DMEM, 10% FBS, 5 μM forskolin (F6886; Sigma), 200 ng/ml recombinant human heregulin-β1 (HRG) (100–03; PeproTech), 10 ng/ml basic fibroblast growth factor (bFGF) (3339-FB: R&D Systems), and 5 ng/ml recombinant rat plateletderived growth factor (PDGF)-AB (1115-AB; R&D Systems); (iii) sustaining dASCs 7d: dASCs were induced for 7 days with complete induction medium; (iv) sustaining dASCs 10d: dASCs were induced for 10 days with complete induction medium; (v) intermittent dASCs: dASCs were induced for 4 days with complete induction medium, after which complete induction medium was replaced with incomplete induction medium consisting of DMEM, 10% FBS, 5 μM forskolin, and 200 ng/ml HRG for induction for 3 days.

Techniques: Derivative Assay, Staining, Flow Cytometry, Modification