polyclonal anti pdgfr β antibodies  (Cell Signaling Technology Inc)

Journal: Neurologia medico-chirurgica

Article Title: Platelet-derived Growth Factor Activates Pericytes in the Microvessels of Chronic Subdural Hematoma Outer Membranes.

doi: 10.2176/jns-nmc.2023-0079

Figure Lengend Snippet: Fig. 1 (A) The outer membranes of chronic subdural hema- tomas (CSDHs) from 8 patients were homogenized in sample buffer and used for western blot analysis with anti-NG2, anti- N-cadherin, anti-VE-cadherin, anti-Tie-2, anti-endothelial ni- tric oxide synthase (eNOS), and anti-β actin antibodies. Nota- bly, all these molecules could be detected in the outer membranes of CSDHs. Positive controls are shown in the right 4 lanes, indicating that these molecules were correctly detect- ed. Liver lysate, rat liver whole cell lysate; NIH/3T3, embryo fi- broblast cell lysate; Endothelial cell, endothelial cell lysate; A375 cell lysate, malignant melanoma cell lysate. (B) Western blot analysis of the same samples was performed with anti-phosphorylated platelet-derived growth factor receptor-β at Tyr751 (p-PDGFR β at Tyr751), anti-PDGFR-β, and anti-β actin antibodies. p-PDGFR β at Tyr751, the activated form of PDGFR-β, was detected. Negative CNT, rat brain whole ly- sate, was used as a negative control; positive CNT, rat spleen whole lysate, was used as a positive control.

Article Snippet: The phosphorylated PDGFR-β immunoblots were stripped and reprobed with primary polyclonal anti-PDGFR-β antibodies (Cell Signaling Technology) at a dilution of 1:750 overnight at 4°C.

Techniques: Western Blot, Derivative Assay, Negative Control, Positive Control

Journal: Neurologia medico-chirurgica

Article Title: Platelet-derived Growth Factor Activates Pericytes in the Microvessels of Chronic Subdural Hematoma Outer Membranes.

doi: 10.2176/jns-nmc.2023-0079

Figure Lengend Snippet: Fig. 2 (A) Hematoxylin and eosin staining demonstrated that the outer membrane included inflammatory cells, fibroblasts, and microvessels. Ten-micrometer slices were immunostained with polyclonal antibodies against platelet-derived growth factor receptor-β (PDGF receptor-β, B and C), N-cadherin (D and E), and Tie-2 (F and G) using the ABC method. The areas within the rect- angle, labeled in B, D, and F, are shown at high magnification in Panels C, E, and G, respectively. Notably, these molecules are ex- pressed in endothelial cells (C, E, and G, arrows). Immunostaining without primary antibodies is shown (H). Scale bars = 50 μm (A, B, D, F, and H).

Article Snippet: The phosphorylated PDGFR-β immunoblots were stripped and reprobed with primary polyclonal anti-PDGFR-β antibodies (Cell Signaling Technology) at a dilution of 1:750 overnight at 4°C.

Techniques: Staining, Membrane, Derivative Assay, Labeling, Immunostaining

Journal: Oncology Letters

Article Title: PDGF-BB regulates the transformation of fibroblasts into cancer-associated fibroblasts via the lncRNA LURAP1L-AS1/LURAP1L/IKK/IκB/NF-κB signaling pathway

doi: 10.3892/ol.2021.12798

Figure Lengend Snippet: PDGF-BB induced the formation of the CAF phenotype in hOMF. (A) The cells were treated with PDGF-BB in a dose-dependent manner (10, 20, 30 ng/ml) in 10% fetal bovine serum for 72 h for three passages. The cells were subjected to western blot analysis with antibodies against CAF markers α-SMA, FAP-α, PDGFR-β and p-PDGFR-β. β-tubulin served as a loading control and sample loading was 20 µg. (B) Fluorescence microscopy of hOMF stained with α-SMA, FAP-α and PDGFR-β (probed with a primary and a secondary antibody). Cells were counterstained with DAPI. Data were expressed as means ± SEM (n=3). *P<0.05 vs. hOMF, # P<0.05 vs. 10 ng/ml PDGF-BB. PDGF, platelet-derived growth factor; CAF, cancer-associated fibroblast; hOMF, human oral mucosa (p3) 500K fibroblast; α-SMA, α-smooth muscle actin; FAP-α, fibroblast activation protein-α; PDGFR-β, platelet-derived growth factor receptor-β; p, phosphorylated.

Article Snippet: The following antibodies were used in the present study: α-SMA (cat. no. ab5694; 1:1,000), FAP-α (cat. no. ab53066; 1:500), IKKα (cat. no. ab32041; 1:1,000), IκBα (cat. no. ab32518; 1:1,000), NF-κB p65 (cat. no. ab16502; 1:1,000), p-NF-κBp65 (p-S536; cat. no. ab86299; 1:1,000), GAPDH (cat. no. ab181602; 1:10,000), β-tubulin (cat. no. ab179511; 1:1,000) and β-actin (cat. no. ab8227; 1:1,000) were obtained from Abcam; LURAP1L (cat. no. PA5-55072; 1:1,000) was obtained from Thermo Fisher Scientific, Inc., and PDGFR-β (cat. no. bs-0232R; 1:1,000) and p-PDGFR-β (Tyr740; cat. no. bs-3323R; 1:1,000) were obtained from BIOSS.

Techniques: Western Blot, Fluorescence, Microscopy, Staining, Derivative Assay, Activation Assay

Journal: Oncology Letters

Article Title: PDGF-BB regulates the transformation of fibroblasts into cancer-associated fibroblasts via the lncRNA LURAP1L-AS1/LURAP1L/IKK/IκB/NF-κB signaling pathway

doi: 10.3892/ol.2021.12798

Figure Lengend Snippet: Effects of the PDGFR-β inhibitor, CP-673451 on LURAP1L-AS1-LURAP1L/IKK/IκB/NF-κB signaling and CAF programming. (A) Reverse transcription-quantitative PCR analysis of LURAP1L and LURAP1L-AS1 expression levels. (B) Western blot analysis of FAP-α, α-SMA, IKKα, IκBα, NF-κB p65 and p-p65 expression following overexpression of LURAP1L-AS1 and treatment with PDGF-BB. *P<0.05 vs. PDGF-BB. PDGF, platelet-derived growth factor; LURAP1L, leucine-rich adaptor protein 1-like; LURAP1L-AS1, LURAP1L antisense RNA 1; IKKα, IκB kinase α; IKK, I-κB kinase; IκBα, nuclear factor of κ light polypeptide gene enhancer in B-cells inhibitor α; NF-κB, nuclear factor-κB; CAF, cancer-associated fibroblasts; p, phosphorylated; α-SMA, α-smooth muscle actin; FAP-α, fibroblast activation protein-α.

Article Snippet: The following antibodies were used in the present study: α-SMA (cat. no. ab5694; 1:1,000), FAP-α (cat. no. ab53066; 1:500), IKKα (cat. no. ab32041; 1:1,000), IκBα (cat. no. ab32518; 1:1,000), NF-κB p65 (cat. no. ab16502; 1:1,000), p-NF-κBp65 (p-S536; cat. no. ab86299; 1:1,000), GAPDH (cat. no. ab181602; 1:10,000), β-tubulin (cat. no. ab179511; 1:1,000) and β-actin (cat. no. ab8227; 1:1,000) were obtained from Abcam; LURAP1L (cat. no. PA5-55072; 1:1,000) was obtained from Thermo Fisher Scientific, Inc., and PDGFR-β (cat. no. bs-0232R; 1:1,000) and p-PDGFR-β (Tyr740; cat. no. bs-3323R; 1:1,000) were obtained from BIOSS.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Over Expression, Derivative Assay, Activation Assay