The following substances were used in this study: Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), and antibiotic–antimycotic agent (Corning Life Science, Corning, NY, USA); sonidegib (Selleckchem, Houston, TX, USA); SP600125 (Calbiochem, Darmstadt, Germany); LPS from Escherichia coli (serotype O111:B4), dimethyl sulfoxide (DMSO), and anti-β-actin and anti-lamin B1 antibodies (Sigma-Aldrich, St. Louis, MO, USA); goat anti-mouse IgG secondary antibody conjugated with Alexa Fluor 488 and 4′,6-diamidino-2-henylindole dihydrochloride (DAPI; Thermo Scientific, Rockford, IL, USA); fluorescent mounting medium (Dako, Carpinteria, CA, USA); MKK4 antibody (Santa Cruz Biotechnology, Dallas, TX, USA); anti-phospho-IκBα antibody (Abcam, Cambridge, MA, USA); and primary antibodies against phospho-MKK4, phospho-JNK, JNK3, phospho-c-Jun, c-Jun, iNOS, COX-2, NF-κB p65, and IκBα and secondary anti-rabbit and anti-mouse antibodies (Cell Signaling Technology, Danvers, MA, USA).
Sonidegib
Manufactured by Selleck Chemicals
Sourced in United States
Sonidegib is a laboratory-grade chemical compound used in various research and development applications. It serves as an important tool for researchers and scientists across diverse fields. The core function of Sonidegib is to facilitate specific experimental procedures and analyses, but its intended use may vary depending on the context of the research being conducted.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum, by Corning (1 mentions)
Fluorescent mounting medium, by Agilent Technologies (1 mentions)
Anti mouse and anti rabbit secondary antibodies, by Cell Signaling Technology (1 mentions)
Antibiotic antimycotic agent, by Corning (1 mentions)
Anti β actin and anti lamin b1 antibodies, by Merck Group (1 mentions)
Dulbecco s modified eagle medium dmem, by Corning (1 mentions)
Gant61, by Selleck Chemicals (1 mentions)
Vismodegib, by Selleck Chemicals (1 mentions)
Tgf β1, by R&D Systems (1 mentions)
Corn oil, by Merck Group (1 mentions)
Tyrphostin ag1024, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
6 protocols using sonidegib
1
Molecular Mechanisms of Anti-inflammatory Signaling
2
Sonidegib Cytotoxicity Assay
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Cells in logarithmic growth phase were grown on 96-well plates (1×104 cells/well). Thereafter, 200 ul sonidegib (Selleck Chemicals, Houston, USA) at concentrations of 0, 5, 10, 20, 30, 40, and 50 μM was added to the wells. Cells of the control group were treated with an equivalent volume of DMSO in place of sonidegib, and a zero-adjustment group was also set up. We then added 10 ul of CCK solution to the cells after 24 h, 48 h, or 72 h, after which the cells were cultured for 2 h at 37°C. The cell culture plate was placed in a microplate reader to read the absorbance at 450 nm. Cell viability (%)=(Absorbance of cells treated with sonidegib-absorbance of zero adjustment)/(Absorbance of control-Absorbance of zero adjustment)×100%.
3
TGFβ1 and Hedgehog Pathway Modulators
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Primary NHBE cells were starved and cultured with basic BEGM medium overnight followed by the treatment with TGFβ1 (R&D Systems, 7754-BH) (5 ng/mL), Hedgehog pathway agonist, SAG (Selleckchem, S6384) (50 nM), an agonist of Smoothened, or various Hedgehog pathway inhibitors, Vismodegib (Selleckchem, S1082) (10 μM), the Smoothened antagonist, Sonidegib (Selleckchem, S2151) (5 μM), and the GLI1 inhibitor, GANT61 (Selleckchem, S8075) (10 μM) for 24 h separately or together as indicated in the figure legends.
4
Hedgehog Pathway Inhibition Assays
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The SMO inhibitor Sonidegib (LDE225) was purchased from Selleckchem. The SHH and IHH blocking antibody MEDI-5304 was kindly provided by MedImmune. Working concentrations were 15 μM for Sonidegib and 30 μg/mL for MEDI-5304. The control plates were treated with equal volumes of vehicle. Cells were pretreated for 48 h before all functional assays. Cells were not treated during sphere- and holoclone-formation assays.
5
Xenograft Models of Malignancies
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All animal procedures were carried out according to animal use protocols approved by the Institutional Animal Care Committee. Immunodeficient NOD-Rag1nullIL2rgnull mice (The Jackson Laboratory) received approximately 3 million cells via subcutaneous flank injection. Cells were in media and Matrigel in a 1:1 ratio. Mice were injected with HSC1λ gGFP (n = 4), HSC1λ gPTCH1 (n = 4), HSC1λ NF1−/− gPTCH1 (n = 4), HSC1λ NF1−/− (n = 12), HSC1λ gGFP (n = 4), HSC1λ gAPC (n = 4). Mice were monitored daily. Mice were sacrificed 4 months post-injection and tumors were harvested. For the sonidegib drug treatment, non-obese diabetic severe combined immune deficiency spontaneous male mice (NOD-SCID-Prkdcscid) received 5 × 106 cells via subcutaneous flank injection. Mice were randomly selected to receive S462TY (10 mice) or STS-26T cells (10 mice). Cells were in a 1:1 media and Matrigel suspension. On day 14, mice were treated with sonidegib (Sellekchem) dissolved in vehicle (PEG 400/5% dextrose in water), at a dose of 20 mg/kg/day, or with vehicle alone. Mice were monitored daily, and tumor size was measured with calipers. Mice were sacrificed when they reached end point (tumor size >1.5 cm in any one dimension).
6
Inhibiting IGF-1 and Hedgehog Pathways
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Some mice (N=3) were administered the IGF-1 inhibitor Tyrphostin AG1024 (Sigma) daily beginning on the day of hydrogel implantation for 21 days. AG1024 (30µg) was delivered in DMSO (3µL) (Sigma) and PBS (200µL) via an IP injection. Vehicle group received DMSO in PBS via an IP injection daily for 21 days. Other mice (N = 3) were administered 10mg/kg/day of Sonidegib (0.25mg; Selleck Chemicals) in 2% DMSO (Sigma) and corn oil (Sigma) for 21 days for the treatment group or 2% DMSO in corn oil for vehicle group via an IP injection. The first dose was administered on the day of hydrogel implantation and continued daily for 21 days. Hydrogels and surrounding tissues were excised and RNA was isolated.
Samples were then assayed for gene expression via RT-qPCR.
Samples were then assayed for gene expression via RT-qPCR.
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