Docetaxel resistant DU145-DR and PC3-DR human PC cells had been previously generated [15 (link)]. Docetaxel, cabazitaxel, cisplatin, carboplatin (MedChemExpress), CXCR2 antagonist SB265610 (Sigma Aldrich) and BCL-2 inhibitor Venetoclax (MedChemExpress) were prepared in DMSO. Western Blot, cell viability, apoptosis and gain and loss of function studies were performed as previously described [16 (link)]. Oligonucleotides and antibodies are listed in Supplementary Table 4 .
Sb265610
Manufactured by Merck Group
Sourced in United States
SB265610 is a laboratory equipment product manufactured by Merck Group. It is designed for use in research and scientific applications. The core function of SB265610 is to facilitate various experimental and analytical procedures, but a detailed description of its intended use is not available.
Automatically generated - may contain errors
Lab products found in correlation
Carboplatin, by MedChemExpress (1 mentions)
Venetoclax, by MedChemExpress (1 mentions)
Gapdh, by Merck Group (1 mentions)
Amd3100, by Merck Group (1 mentions)
Ip 10, by R&D Systems (1 mentions)
Sdf 1α, by R&D Systems (1 mentions)
Matrigel, by BD (1 mentions)
Mtesr1 complete kit, by STEMCELL (1 mentions)
Transwell polycarbonate permeable supports, by Merck Group (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Gm csf, by Thermo Fisher Scientific (1 mentions)
Y 27632, by Merck Group (1 mentions)
Fludarabine, by Merck Group (1 mentions)
5 15 dpp, by Merck Group (1 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Ix71 microscope, by Olympus (1 mentions)
Matrigel, by Corning (1 mentions)
Pgf2α, by Cayman Chemical (1 mentions)
Mab208, by R&D Systems (1 mentions)
10 protocols using sb265610
1
Characterizing Docetaxel-Resistant Prostate Cancer Cells
2
Overexpression of S100A8/A9 Proteins
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To achieve overexpression, we designed plasmids carrying full-length cDNAs of S100A8 and S100A9 from mouse and human sources, respectively. Through DNA sequencing, we ensured that the plasmids, namely pIRES-mS100A8, pIRES-mS100A9, pEGFP-hS100A8, and pEGFP-hS100A8, precisely matched the base sequences of their respective cDNAs. The following inhibitors were utilized in the experiments: pD098059 (ERK inhibitor, #S1805) was purchased from Kangwei Century; BAY11-7082 (NF-κB inhibitor, #S1523) was purchased from Byotime; AG490 (STAT3 inhibitor, #S1143) was purchased from Selleck; SB265610 (CXCR2 inhibitor, #SML0421) was purchased from SIGMA; Primary antibodies: NF-κB P65(C22B4) Rabbit mAb (#4764S), Phospho-IKKα/β (Ser176/180) Rabbit mAb (#2697), p44/42 MAPK (Erk1/2) Rabbit mAb (#4695) , Phospho-p44/42 ( Thr202/Tyr204) Antibody (#9101) , Phospho-Stat3 (Tyr705) Rabbit mAb (#9145) purchased from CST; Rabbit Anti-CXCR2 antibody (#ab14935), Rabbit Anti-CXCL5 antibody(#ab14935), Rabbit Anti-CXCR2 antibody(#ab14935), Rabbit Anti-CXCL5 antibody(#ab9983) were purchased from Abcam; secondary antibody: GAPDH Mouse monoclonal antibody(#MAB3740) was purchased from MILLIPORE.
3
Chemokine Receptor Modulation in hiPSCs
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The p27 hiPSC cell line hNF C11 was purchased from the Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences. Both newly-established hESCs and hiPSCs were seeded in 6-well plates pre-coated with 1% Matrigel (BD Biosciences) and cultured with the mTeSR1 complete kit (Stem Cell Technology, USA). Recombinant human chemokine proteins IL-8, SDF-1α, and IP-10 were purchased from R&D Systems (USA). CXCR1/CXCR2-specific antagonist reparixin and CXCR3-specific antagonist NBI74330 were bought from MCE (USA). CXCR4-specific antagonist AMD3100 and CXCR2-specific antagonist SB265610 were purchased from Sigma (USA). The hPSCs were treated by either 1 μg/ml of IL-8, 100 nM reparixin, 100 nM SB265610, 100 ng/ml of SDF-1α, 100 nM AMD3100, 50 ng/ml of IP-10, or 100 nM NBI74330 overnight.
4
MDSC Differentiation and Migration Assay
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MDSCs were cultured as previously described.30 (link) Briefly, BM cells were cultured in the presence of 10 ng/mL GM-CSF (Peprotech, Rocky Hill, NJ) and 10 ng/mL IL-4 (Peprotech, Rocky Hill, NJ) for 4 days to induce the differentiation to MDSCs. Supernatants from control or IL-1α-overexpressing hepa1-6 cells were added into 24-well plates with Transwell polycarbonate-permeable supports (pore size = 8.0 μm; Merck Millipore, Burlington, MA). 2 × 105 MDSCs were preincubated with or without CXCR2 antagonist, SB265610 (Sigma, St. Louis, MO), for 30 min and seeded on the upper chambers of the inserts. After incubation for 18 hours, the MDSCs in the bottom compartment were counted.
5
SB-265610 Intraperitoneal Injection
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6
Protein Assay Reagents Protocol
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Y-27632, fludarabine, 5,15 DPP and SB-265610 were obtained from Sigma (St. Louis, MO, USA). Recombinant HIV-p17 protein was provided by Medestea (Torino, Italy).
7
Endothelial Cell Proliferation Assay
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HRMECs were spread into 96‐well plates at a concentration of 2 × 105 cells/ml, and PGF2α [20–2,500 nM (Goupil et al, 2010 (link)), Cayman, 16010], SB265610 (1 μM, Sigma, SML0421), or equal volumes of DMSO were added to each well, depending on the treatment group. Cells were then incubated at 37°C for 24 h. Cell proliferation was assessed using the Cell Counting Kit‐8 (Dojindo, CK04), according to the manufacturer's instructions.
8
Targeting CXCR2 in Hepatic Tumor Model
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Mice were injected intrahepatically with hepa1-6 cells stably expressing secreted IL-1α. One day after tumor injection, mice were randomly divided into two groups and were treated with CXCR2 inhibitor SB265610 (Sigma, St. Louis, MO) at a dose of 2 mg/kg mouse body weight every 3 days. Control group mice were treated with equal volume of DMSO (Sigma, St. Louis, MO).
9
Endothelial Cell Tube Formation
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HRMECs were spread at a density of 1 × 105 cells per well in 24‐well plates coated with low growth factor Matrigel (Corning, 356230). PGF2α (500 nM, Cayman, 16010), SB265610 (1 μM, Sigma, SML0421), or equal volumes of DMSO were added to each well, depending on the treatment group. Cells were then incubated at 37°C for 12 h. Cell tube formation in each well was photographed using an IX71 microscope (Olympus). The total tubule length, number of junctions, number of meshes, and percentage of mesh area were analyzed using the Angiogenesis Analyzer plugin in ImageJ.
10
Ovarian Cancer Cell Line Maintenance
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The HIO-80 cell line, an immortalized human ovarian epithelial cell line, was obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA) and maintained in a 1:1 mixture of medium 199 and MCDB-105 (Sigma-Aldrich; St. Louis, MO, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Thermo Fisher Scientific; Waltham, MA, USA) and 0.2 units/mL of insulin (Sigma-Aldrich). The SKOV3 (clear cell) and OVCAR3 (high-grade serous ovarian cancer, HGSOC) cell lines were also obtained from ATCC and maintained in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% FBS.
The Human CXCL8 neutralizing antibody, MAB208, was purchased from R&D systems (Minneapolis, MN, USA). Selective CXCR2 antagonists, SB225002 and SB265610, were obtained from Sigma-Aldrich.
The Human CXCL8 neutralizing antibody, MAB208, was purchased from R&D systems (Minneapolis, MN, USA). Selective CXCR2 antagonists, SB225002 and SB265610, were obtained from Sigma-Aldrich.
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