Journal: Clinical & Translational Immunology

Article Title: Distinct macrophage phenotypes skewed by local granulocyte macrophage colony‐stimulating factor (GM‐CSF) and macrophage colony‐stimulating factor (M‐CSF) are associated with tissue destruction and intimal hyperplasia in giant cell arteritis

doi: 10.1002/cti2.1164

Figure Lengend Snippet: FRβ + positivity in the inner intima is associated with high‐degree intimal hyperplasia. Classification of mildly and massively occluded TAB based on the thickness of intima (a) . The Mann–Whitney U ‐test showed significantly higher expression of FRβ in the inner intima of TABs with massive intimal hyperplasia (mild occlusion n = 6; massive occlusion n = 5) (b) . The Wilcoxon signed‐rank test showed higher expression of PDGF‐AA in M‐MØ compared to GM‐MØ ( n = 10 each, ELISA was performed in duplicates for each sample) (c) . GCA, giant cell arteritis; GM‐MØ, GM‐CSF macrophages; M‐MØ, M‐CSF macrophages; TAB, temporal artery biopsy.

Article Snippet: PDGF‐AA concentrations in the supernatant of activated cultures were measured with the Human PDGF‐AA Duo set ELISA (DY‐221, R&D Systems) according to the manufacturer's instruction.

Techniques: MANN-WHITNEY, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Circulation

Article Title: Mechanosensitive p27 Kip1 Regulation and Cell Cycle Entry in Vascular Smooth Muscle Cells

doi: 10.1161/01.cir.0000079102.08464.e2

Figure Lengend Snippet: Figure 6. Stretch-induced Akt activation is growth factor and growth factor receptor independent. a, Neutralizing antibodies (nAb) against IGF, bFGF, or PDGF were added, and cells were stretched for 15 minutes. Phosphorylation of Akt was determined by immunoblotting (WB) with phosphospecific antibody. Pan-Akt served as housekeeping protein. b, Growth factor receptor inhibitors AG1478 (EGF), AG1296 (PDGF), and AG1024 (IGF-1) were added, and cells were stretched for 15 minutes or 24 hours to determine Akt activation and p27Kip1 expression by immunoblotting (WB), respectively. Pan-Akt served as housekeeping protein.

Article Snippet: Mouse monoclonal anti-p27Kip1, antiretinoblastoma (RB), anti-Cdk2 (Santa Cruz Biotechnology, Santa Cruz, Calif), anti-basic fibroblast growth factor (bFGF), antiinsulinlike growth factor (IGF), and anti–platelet-derived growth factor (PDGF) (R&D Systems, GMBH, Weisbaden, Germany) were also used.

Techniques: Activation Assay, Phospho-proteomics, Western Blot, Expressing

Journal: International journal of oncology

Article Title: Synergistic effects of imatinib and carboplatin on VEGF, PDGF and PDGF-Rα/ß expression in squamous cell carcinoma of the head and neck in vitro.

doi: 10.3892/ijo.2011.912

Figure Lengend Snippet: Figure 1. Immunohistochemistry for PDGF-α expression; (A and B) Positive immunohistochemical reactivity against PDGF-α (x20) after incubation with carboplatin and imatinib (3+18 µmol) after 48 h. (C and D) Positive immunohistochemical reactivity against PDGF-α (x20-40) with carboplatin and imatinib (7.5+30 µmol) after 120 h. (E-F) Control group with strong immunoreactivity.

Article Snippet: Immunohistochemistry of PDGF-A/B ligand. immunohistochemical studies were performed using a monoclonal rabbit antibody directed against human PDGF-A/B (ACRIS Antibodies, Herford, Germany).

Techniques: Immunohistochemistry, Expressing, Immunohistochemical staining, Incubation, Control

Journal: bioRxiv

Article Title: A non-canonical role and regulations of polo-like kinase-4 in fibroblast cell-type transition

doi: 10.1101/570267

Figure Lengend Snippet: Rat primary adventitial fibroblasts were cultured in the complete medium, starved in the basal medium overnight (see Methods), and pretreated for 2h with vehicle (equal amount of DMSO) or the PLK4-selective inhibitor centrinone-B (Cen-B) at indicated concentrations, followed by stimulation with 60 ng/ml PDGF-AA (abbreviated as AA throughout). Cells were harvested at 24h after stimulation (or otherwise specifically indicated) for various assays. A. Morphology. Cells were (or not) stimulated by AA for 24h without or with pre-treatment (1µM Cen-B). Green fluorescent calcein was used to illuminate cell morphologies. B. Proliferation. CellTiterGlo assay was performed after 72h AA stimulation of cells without or with pretreatment by Cen-B at increasing concentrations. C. Migration (scratch assay). Cells were (or not) stimulated by AA for 24h without or with pre-treatment (1 or 10 µM Cen-B). Calcein was used to illuminate the cells. D. Western blots of αSMA and vimentin. Protein band densitometry was normalized to loading control (β-actin), and then to the basal condition (DMSO, no AA), and finally quantified as fold change. Fold changes from at least 3 independent experiments were averaged, and mean ± SEM was calculated. Quantification for C and D: Mean ± SEM, n ≥3 experiments. One-way ANOVA/Bonferroni test: ***P< 0.001 compared to the condition of AA without Cen-B. E. Effect of PLK4 silencing. Cells were transfected with scrambled or PLK4-specific siRNA for 48 h in the complete medium, starved overnight, and then stimulated with AA for 10 min before harvest for Western blotting. F. Effect of PLK4 inhibition on MRTF-A. Cells were pretreated with 1µM Cen-B (or vehicle) and then stimulated (or not) with AA for 48 h. G. Luciferase reporter assay of SRF transcriptional activity. Cells were transfected with the empty vector control or the SRF-Luciferase vector, followed by luminescence reading. The condition with 5 µM tubastatin-A, an HDAC6 inhibitor and a novel SRF stimulator , served as a positive control. Mean ± SEM, n = 3 experiments. Student’s t-test: **P <0.01, ***P<0.001, between two gray bars.

Article Snippet: For induction of fibroblast cell-type transition, cells were first starved overnight in Fibroblast Basal Medium (Cat. 2267b, Cell Biologics Inc.) that contains no FBS, and then stimulated with 60 ng/ml PDGF-AA (rat recombinant, R&D Systems Inc., MN) for specifically indicated (in figures) length of time.

Techniques: Cell Culture, Migration, Wound Healing Assay, Western Blot, Transfection, Inhibition, Luciferase, Reporter Assay, Activity Assay, Plasmid Preparation, Positive Control

Journal: Biology of reproduction

Article Title: Proliferation and differentiation of rat theca-interstitial cells: comparison of effects induced by platelet-derived growth factor and insulin-like growth factor-I.

doi: 10.1095/biolreprod60.3.546

Figure Lengend Snippet: FIG. 2. Effects of PDGF (0.1–1 nM) on DNA synthesis by rat T-I cells in the presence and in the absence of IGF-I (10 nM). The cells were cultured in serum-free medium for 48 h in 96-well plates at a concentration of 35 000 cells/well. Each well contained 0.25 ml of medium. During the last 24 h of culture, [3H]thymidine (1 mCi/well) was added to determine DNA synthesis. Bars indicate the mean of each treatment, and vertical lines indicate the SEM from at least six replicates; means with no superscripts in common are significantly different (p , 0.05).

Article Snippet: Recombinant PDGF-AB heterodimer and anti-human PDGF neutralizing antibody (Ab-PDGF) were purchased from R&D Systems (Minneapolis, MN).

Techniques: DNA Synthesis, Cell Culture, Concentration Assay

Journal: Biology of reproduction

Article Title: Proliferation and differentiation of rat theca-interstitial cells: comparison of effects induced by platelet-derived growth factor and insulin-like growth factor-I.

doi: 10.1095/biolreprod60.3.546

Figure Lengend Snippet: FIG. 3. Effect of Ab-PDGF (10 mg/ml) on DNA synthesis by rat T-I cells. The cells were cultured in serum-free medium with or without PDGF (1 nM), IGF-I (10 nM), and/or Ab-PDGF (10 mg/ml) for 48 h. The cultures were carried out in 96-well plates at a concentration of 35 000 cells/well. Each well contained 0.25 ml of medium. During the last 24 h of culture, [3H]thymidine (1 mCi/well) was added to determine DNA synthesis. Bars indicate the mean of each treatment, and vertical lines indicate the SEM from at least six replicates; * denotes values significantly different from control (p , 0.05).

Article Snippet: Recombinant PDGF-AB heterodimer and anti-human PDGF neutralizing antibody (Ab-PDGF) were purchased from R&D Systems (Minneapolis, MN).

Techniques: DNA Synthesis, Cell Culture, Concentration Assay, Control

Journal: Biology of reproduction

Article Title: Proliferation and differentiation of rat theca-interstitial cells: comparison of effects induced by platelet-derived growth factor and insulin-like growth factor-I.

doi: 10.1095/biolreprod60.3.546

Figure Lengend Snippet: FIG. 4. Effect of PDGF (1 nM) and IGF-I (10 nM) on the number of steroidogenically active [3b-HSD(1)] and steroidogenically inactive [3b- HSD(2)] rat T-I cells. T-I cells were cultured in serum-free medium with or without treatments for 48 h in 24-well plates at a concentration of 350 000 cells per well. Each well contained 1 ml of medium. At the end of the culture period the cells were trypsinized, and steroidogenically active cells were stained by a histochemical reaction identifying 3b-HSD activ- ity. Bars indicate the mean for each treatment, and vertical lines indicate the SEM from four cultures; * denotes values significantly different from control; p , 0.05.

Article Snippet: Recombinant PDGF-AB heterodimer and anti-human PDGF neutralizing antibody (Ab-PDGF) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Cell Culture, Concentration Assay, Staining, Control