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Nvp lde225

Manufactured by Selleck Chemicals
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Sourced in Germany

NVP-LDE225 is a laboratory compound used for research purposes. It is a small molecule that acts as an inhibitor, though its specific function and intended applications are not provided in this factual, unbiased description.

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Lab products found in correlation

Gdc 0449, by Selleck Chemicals (2 mentions) Gammacell 40 exactor, by Best Theratronics (2 mentions) Cx 4945, by Selleck Chemicals (1 mentions) Mtt kit, by Beyotime (1 mentions) Anti β actin antibody, by Merck Group (1 mentions) Anti flag antibody f3165, by Merck Group (1 mentions) Mk 2206, by Selleck Chemicals (1 mentions) Paclitaxel, by Selleck Chemicals (1 mentions) Bevacizumab, by Roche (1 mentions) Everolimus, by Selleck Chemicals (1 mentions) Sunitinib, by Selleck Chemicals (1 mentions) Gant61, by Selleck Chemicals (1 mentions) Tubulin, by Merck Group (1 mentions) Rad001, by Selleck Chemicals (1 mentions) Gant58, by Bio-Techne (1 mentions) Prmt1, by Cell Signaling Technology (1 mentions) Ami 1, by Merck Group (1 mentions) Sybr green real time pcr kit, by Bio-Rad (1 mentions) Gant61, by Bio-Techne (1 mentions) Flag m2 magnetic beads, by Merck Group (1 mentions)

6 protocols using nvp lde225

1

Lentiviral Expression of SMO Variants

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SMOWT was purchased from Sino Biological (https://www.sinobiological.com/) and SmoM2 (W535L)‐pcw107‐V5 plasmids were purchased from Addgene (http://www.addgene.org/). The SMO expression frame was subcloned into pCDH‐CMV‐MCS‐FE1a‐Puro vector (System Biosciences) with a Flag‐tag at the C‐terminal of SMO. The constructed plasmids were used to package lentivirus in 293T cells according to Kutner's protocol.34 Anti‐Flag antibody (#F3165) and anti‐β‐actin antibody (#A5441) were from Sigma Aldrich. MTT kit (#C0009S) was purchased from Beyotime (https://beyotime.com/). SAG (SMO agonist) (#S7779) (0.1 µM), GDC‐0449 (#S1082) (1 µM), NVP‐LDE225 (#S2151) (1 µM), CX4945 (#S0707), and MK2206 (#S1078) were purchased from Selleck (https://www.selleck.cn/).
Yao Y., Wang Y., Yang F., Wang C., Mao M., Gai Q., He J., Qin Y., Yao X., Lan X., Zhu J., Lu H., Zeng H., Yao X., Bian X, & Wang Y. (2022). Targeting AKT and CK2 represents a novel therapeutic strategy for SMO constitutive activation‐driven medulloblastoma. CNS Neuroscience & Therapeutics, 28(7), 1033-1044.
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2

Tumor Cell Proliferation Assays

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To measure effects of inhibitors in combination with radiation, tumor cells were plated in 96-well plates at a density of 0.2 × 106 cells per well and cultured in the presence of DMSO, 50 nM YM155, or 10 ug/ml S12. After 24hrs, cells were subjected to 0, 0.25, or 0.5 Gy radiation using a Gammacell 40 Exactor (Low-dose cesium 137 irradiator, Best Theratronics Ltd., Ottawa, Ontario, Canada). Cells were cultured for an additional 24 hr, and [methyl-3H]thymidine assays were performed as described above.
To measure effects of inhibitors in combination with the SHH antagonist NVP-LDE225 (Selleck Chemicals, S2151), tumor cells were plated in 96 well plates at 0.2 × 106 cells/well and cultured with increasing doses of LDE225 or a single dose of Survivin antagonist (10μg/ml S12, 20nM YM255) alone or in combination with LDE225 as indicated. Cells were cultured for 48hrs and [methyl-3H]thymidine assays were performed as described above.
Brun S.N., Markant S.L., Esparza L.A., Garcia G., Terry D., Huang J.M., Pavlyukov M.S., Li X.N., Grant G.A., Crawford J.R., Levy M.L., Conway E.M., Smith L.H., Nakano I., Berezov A., Greene M.I., Wang Q, & Wechsler-Reya R.J. (2014). Survivin as a therapeutic target in Sonic hedgehog-driven medulloblastoma. Oncogene, 34(29), 3770-3779.
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3

Breast Cancer Cell Line Characterization

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NVP-LDE225 and paclitaxel were purchased from Selleck Chemicals (Munich, Germany). Bevacizumab was kindly provided by Roche (Basel, Switzerland).
For this study, a panel of immortalised (MDA-MB-361, SKBR-3, MCF7, MDA-MB-231, HCC70 and MDA-MB-468, KPL-4, JIMT-1) and primary (K90, K79, K193 and K197) breast cancer cell lines were used. Besides, some experiments have been performed on endothelial cells such as human umbilical vein endothelial cells (HUVECs) and mouse brain endothelial cell 5 (bEND5). For a detailed description, please see the Supplementary Methods section.
Di Mauro C., Rosa R., D'Amato V., Ciciola P., Servetto A., Marciano R., Orsini R.C., Formisano L., De Falco S., Cicatiello V., Di Bonito M., Cantile M., Collina F., Chambery A., Veneziani B.M., De Placido S, & Bianco R. (2017). Hedgehog signalling pathway orchestrates angiogenesis in triple-negative breast cancers. British Journal of Cancer, 116(11), 1425-1435.
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4

Anticancer Drug Compounds Evaluation

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NVP-LDE225, everolimus, sunitinib, and GANT-61 were purchased from Selleck Chemicals, Munich, Germany.
D'Amato C., Rosa R., Marciano R., D'Amato V., Formisano L., Nappi L., Raimondo L., Di Mauro C., Servetto A., Fulciniti F., Cipolletta A., Bianco C., Ciardiello F., Veneziani B.M., De Placido S, & Bianco R. (2014). Inhibition of Hedgehog signalling by NVP-LDE225 (Erismodegib) interferes with growth and invasion of human renal cell carcinoma cells. British Journal of Cancer, 111(6), 1168-1179.
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5

Tumor Cell Proliferation Assays

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To measure effects of inhibitors in combination with radiation, tumor cells were plated in 96-well plates at a density of 0.2 × 106 cells per well and cultured in the presence of DMSO, 50 nM YM155, or 10 ug/ml S12. After 24hrs, cells were subjected to 0, 0.25, or 0.5 Gy radiation using a Gammacell 40 Exactor (Low-dose cesium 137 irradiator, Best Theratronics Ltd., Ottawa, Ontario, Canada). Cells were cultured for an additional 24 hr, and [methyl-3H]thymidine assays were performed as described above.
To measure effects of inhibitors in combination with the SHH antagonist NVP-LDE225 (Selleck Chemicals, S2151), tumor cells were plated in 96 well plates at 0.2 × 106 cells/well and cultured with increasing doses of LDE225 or a single dose of Survivin antagonist (10μg/ml S12, 20nM YM255) alone or in combination with LDE225 as indicated. Cells were cultured for 48hrs and [methyl-3H]thymidine assays were performed as described above.
Brun S.N., Markant S.L., Esparza L.A., Garcia G., Terry D., Huang J.M., Pavlyukov M.S., Li X.N., Grant G.A., Crawford J.R., Levy M.L., Conway E.M., Smith L.H., Nakano I., Berezov A., Greene M.I., Wang Q, & Wechsler-Reya R.J. (2014). Survivin as a therapeutic target in Sonic hedgehog-driven medulloblastoma. Oncogene, 34(29), 3770-3779.
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6

Antibody Specifications and Peptide Details for Methylated Gli1 Analysis

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The antibodies used in this study were Gli1 (#3538, Cell Signaling Technology, for western blotting and chromatin immunoprecipitation (ChIP) assay, and#sc-20687, Santa Cruz Biotechnology, for immunohistochemistry), actin (#A2066, Sigma-Aldrich), Flag (#F3165, Sigma-Aldrich), Flag M2 magnetic beads (#M8823, Sigma-Aldrich), PRMT1 (#2449, Cell Signaling Technology), and tubulin (#T5168, Sigma-Aldrich). GDC-0449, NVP-LDE225, and RAD-001 were purchased from Selleck Chemicals LLC, GANT58 and GANT61 from Tocris Bioscience, and AMI-1 from Sigma-Aldrich. The SYBR Green real-time PCR kit was obtained from Bio-Rad and TGF-β from Peprotech (#100-21). The antibody against R597-methylated Gli1 was developed using the following synthetic peptide with asymmetric dimethylation at R597 as antigen: RARYASA-[R597(aMe2)]-GGGTS (C-terminal amidation and N-terminal KLH conjugation). Other peptides used in the study included the following:

Cold Gli1 peptide without R597 methylation [Gli1-R597]: RARYASA-[R597]-GGGTS;

Hot Gli1 peptide with R597 monomethylation [Gli1-R597(Me)]: RARYASA-[R597(Me)] -GGGTS;

Hot Gli1 peptide with R597 asymmetric dimethylation [Gli1-R597(aMe2)]: RARYASA-[R597(aMe2)]-GGGTS;

Hot Gli1 peptide with R597 symmetric dimethylation [Gli1-R597(sMe2)]: RARYASA-[R597(sMe2)]-GGGTS.

Wang Y., Hsu J.M., Kang Y., Wei Y., Lee P.C., Chang S.J., Hsu Y.H., Hsu J.L., Wang H.L., Chang W.C., Li C.W., Liao H.W., Chang S.S., Xia W., Ko H.W., Chou C.K., Fleming J.B., Wang H., Hwang R.F., Chen Y., Qin J, & Hung M.C. (2016). Oncogenic functions of Gli in pancreatic adenocarcinoma are supported by its PRMT1-mediated methylation. Cancer research, 76(23), 7049-7058.
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