Bevacizumab, an immunoglobulin (Ig) G1 monoclonal antibody targeting VEGF-A, was obtained from Chugai Pharmaceutical (Tokyo, Japan). For in vivo studies, bevacizumab was diluted in phosphate-buffered saline (PBS). BGJ-398, a FGFR-specific inhibitor, was purchased from Chemietek (Indianapolis, IN). For the in vitro studies, 4 mM stock solutions were prepared in DMSO. For the in vivo studies, BGJ-398 was formulated in PEG300/D5W (2:1). FGF2-neutralizing antibody was purchased from R&D systems (Minneapolis, MN). AMD3100, a selective CXCR4 antagonist, was purchased from Sigma (St Louis, MO). For in vitro studies, 10 mg ml−1 stock solutions were prepared in PBS. SU5416, a VEGFR-specific inhibitor, was purchased from Abcam (Cambridge, MA). For the in vivo studies, SU5416 was formulated in DMSO. An anti-mouse IL-2 receptor β-chain monoclonal antibody, TM-β1, was supplied by Drs M. Miyasaka and T. Tanaka (Osaka University, Osaka, Japan).
Su5416
Manufactured by Abcam
Sourced in United Kingdom
SU5416 is a potent, ATP-competitive inhibitor of the receptor tyrosine kinase VEGFR2 (Flk-1/KDR). It inhibits VEGF-induced autophosphorylation of VEGFR2 and subsequent downstream signaling pathways.
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Lab products found in correlation
5 protocols using su5416
1
Targeting VEGF and FGF Signaling
2
Hypoxia-Induced Pulmonary Hypertension in Rats
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Male rats weighing 90 to 110 g (CLEA) were used in our experiments. The rat PH model was generated as described previously [12 (link),13 (link)]. In brief, 20 mg/kg of vascular endothelial growth factor inhibitor SU5416 (Abcam) suspended in CMC (0.5% [w/v] carboxymethylcellulose sodium, 0.9% [w/v] sodium chloride, 0.4% [v/v] polysorbate 80, and 0.9% [v/v] benzyl alcohol in deionized water) was subcutaneously injected at Day 0. The rats were housed in a hypoxic chamber (10% O2) maintained using a hypoxic air generator (TEIJIN) and monitored with an O2 analyzer (JIKO-255) for 3 weeks, and then re-exposed to normoxia for an additional 3 or 5 weeks. A purified AIN-93G containing 1,000 IU/kg of cholecalciferol (normal diet) and the modified AIN-93G containing 10,000 IU/kg of that (high vitamin D diet) were purchased from Oriental Yeast Co., Ltd (Japan). PH rats were divided into two groups and fed different dietary amounts of cholecalciferol on ad libitum.
3
Inhibitors of Angiogenesis and Fibrosis
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Nintedanib and SB431542 were obtained from Boehringer Ingelheim GmbH & Co. KG (Biberach, Germany). SU5416, a VEGFR-specific inhibitor, was purchased from Abcam (Cambridge, MA). BGJ-398 and imatinib were purchased from Chemietek (Indianapolis, IN). Bleomycin (BLM) was purchased from Nippon Kayaku Co. (Tokyo, Japan).
4
Rat Emphysema Model and MSC Therapy
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The experimental procedures were conducted under EC Directive 86/609/EEC for animal experiments and approved by the Institutional Animal Care and Use Committee of Jiangsu University (No. UJS-IACUC-2019040901). Our experiments conformed to the effective laws and ethical recommendations currently in China. Male Sprague-Dawley (SD) rats were purchased from Jiangsu University and kept in the Laboratory Animal Research Center of Jiangsu University. Emphysema models were established using the VEGF receptor blocker, SU5416 as described previously (17 (link)). Rats were divided randomly into four groups: (A) Control group (CON, n=9), (B) Control＋MSC group (CON＋MSC, n=9), (C) SU5416 (SU, n=9) and (D) SU5416＋MSC group (SU＋MSC, n=9). Rats in group C and D were administered with SU5416 (Abcam, Cambridge, UK) subcutaneously at a dosage of 20 mg/kg body weight, three times a week for 3 weeks. SU5416 was dissolved in diluents containing 0.5% carboxymethyl-cellulose sodium, 0.9% sodium chloride, 0.4% polysorbate 80, and 0.9% benzyl alcohol in deionized water (17 (link)). Rats in group A and B only received the diluents administered subcutaneously. At the end of the second week after administration of SU5416 or diluents, 3×106 hUC-MSCs were administered into groups B and D, at the third passage resuspended in 0.3 ml PBS through the tail vein, whereas groups A and C received only 0.3 ml PBS.
5
Investigating Intraventricular Hemorrhage Pathways
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Briefly, this study has three parts. First, rats were randomly divided into two groups (sham, IVH). The IVH group received an injection of 200 μL homologous blood while the sham group received the same volume of saline into the right lateral ventricle. Rats were euthanized at day 7, the brains were used for ventricular volume calculation (n = 6 per group), western blot (n = 6 per group) and brain histology (n = 4 per group). Second, rats were treated with metformin [65 (link),66 (link)] (Abcam, 50 mg/kg, i.p.) or saline (equal volume, i.p.) daily for the following 7 days after IVH, then were euthanized at Day 7. The brains were used for ventricular volume calculation (n = 6 per group), western blot (n = 6 per group), internalization assay (n = 6 per group) and brain histology (n = 4 per group). Third, rats were divided into sham, IVH + Vehicle (2% dimethyl sulfoxide in saline), IVH + VEGFR2 inhibitor (SU5416, MCE, 25 mg/kg, i.p.) and IVH + p-Src inhibitor (PP2, Abcam, 1 mg/kg, i.p.). SU5416 or PP2 were injected at day 0 and day 3 [13 (link),67 (link),68 (link)]. At day 7, rats were euthanized and the brains were used for ventricular volume calculation (n = 6 per groups), western blot (n = 6 per groups) and brain histology (n = 4 per groups) (Figure 6 ).
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