Blz945

Manufactured by Selleck Chemicals

BLZ945 is a laboratory equipment designed for chemical analysis and synthesis. It is a compact and versatile instrument that can perform various tasks in a research or industrial setting. The core function of BLZ945 is to facilitate precise measurement, mixing, and reaction monitoring of chemical substances. This equipment is suitable for use in a wide range of applications, including but not limited to, organic synthesis, analytical chemistry, and material science research.

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Lab products found in correlation

8 protocols using blz945

Screening Novel Kinase Inhibitors in Ba/F3 Cells

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Pexidartinib and BLZ945 were purchased from Selleckchem. LOXO-292 (Selpercatinib) was purchased from Active Biohem. Drug studies were conducted in vitro using fluorescence-activated cell-sorted DAPI⁻eGFP⁺ Ba/F3 cells that stably expressed the MIGII-empty vector, MIGII-CSF-1R (Y546_K551del), MIGII-CSF-1R (W450_E456del), CSF-1R (Y561_I564del), and CSF-1R (G936S) and MIGII-NCOA4-RET constructs using the CellTiter-Glo Luminescent Cell Viability Assay from Promega Corporation, according to the manufacturer’s instructions. The MIGII CSF-1R (Y546_K551del), MIGII-CSF-1R (W450_E456del), CSF-1R (Y561_I564del), and CSF-1R (G936S) and MIGII-NCOA4-RET, fluorescence-activated cell-sorted Ba/F3 cells were maintained in RPMI + 10% FBS + penicillin and streptomycin medium, without mouse IL-3. MIGII-EV was maintained in RPMI + 10% FBS + penicillin and streptomycin with recombinant mouse IL-3 (1 ng/ml).

Durham B.H., Rodrigo E.L., Picarsic J., Abramson D., Rotemberg V., De Munck S., Pannecoucke E., Lu S.X., Pastore A., Yoshimi A., Mandelker D., Ceyhan-Birsoy O., Ulaner G.A., Walsh M., Yabe M., Petrova-Drus K., Arcila M.E., Ladanyi M., Solit D.B., Berger M.F., Hyman D.M., Lacouture M.E., Erickson C., Saganty R., Ki M., Dunkel I.J., López V.S., Mora J., Haroche J., Emile J.F., Decaux O., Geissmann F., Savvides S.N., Drilon A., Diamond E.L, & Abdel-Wahab O. (2019). Activating mutations in CSF-1R and additional receptor tyrosine kinases in histiocytic neoplasms. Nature medicine, 25(12), 1839-1842.

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Screening Novel Kinase Inhibitors in Ba/F3 Cells

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Characterizing Human Cancer Cell Lines

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The human colorectal cancer cell line HCT116 (ATCC), the human breast cancer cell lines MCF-7 (ATCC), and the human monocyte cell lines THP-1 were maintained RPMI-1640 medium (Gibco) supplemented with 10% (v/v) fetal bovine serum (Biological Industries). All cells were grown in a humidified incubator (Thermo Scientific) containing 5% CO₂ at 37 °C, and had not been passaged for 3 months before the experiment. The cell line was routinely tested to exclude mycoplasma contamination and characterized by the use of short tandem repeat markers by the Genetic Testing Biotechnology Company (Suzhou, China). All siRNA oligos and plasmids were introduced into cells by Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instruction. Use 1 μg/mL doxorubicin (Selleck Chemicals) and DMSO as a control to perform the specified analysis on the cells. For drug treatments, HCT116 and MCF-7 cells were pulse-exposed to doxorubicin for 2 h, and then replaced with fresh medium, and the cells were cultured for additional several days. THP-1 cells were differentiated into macrophages by treatment with 320 nM of phorbol 12-myristate 13-acetate (PMA) for 24 h, and then recovered 2/3 of conditioned medium and 1/3 of fresh medium. THP-1 cells were treated with 500 nM BLZ945 (Selleck Chemicals) for 48 h to inhibit M-CSF/M-CSFR axis.

Liu H., Zhao D., Li H., Zhang W., Lin Q., Wang X., Zheng S., Zhang L., Li L., Hu S, & Hu Y. (2022). Blocking iASPP/Nrf2/M-CSF axis improves anti-cancer effect of chemotherapy-induced senescence by attenuating M2 polarization. Cell Death & Disease, 13(2), 166.

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Genetic and Pharmacological Inhibition of CSF1R

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For genetic depletion of CSF1 receptor, we gave doxycycline (SIGMA, 20 mg/mL) in drinking water with 5% w/v sucrose to the rtTA:tetO-Cre:Csf1r^F/F (Csf1r cKO) mice from 7 days after tumor injection to the endpoint. For pharmacological CSF1R inhibition, we gave BLZ945 (Selleckchem, 4 mg/20 g mouse, oral gavage) from 7 days after tumor injection to the endpoint.

Kitamura T., Doughty-Shenton D., Cassetta L., Fragkogianni S., Brownlie D., Kato Y., Carragher N, & Pollard J.W. (2018). Monocytes Differentiate to Immune Suppressive Precursors of Metastasis-Associated Macrophages in Mouse Models of Metastatic Breast Cancer. Frontiers in Immunology, 8, 2004.

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Transwell Assay for Monocyte-Derived Langerhans Cell Migration

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Migration of mo-LCs was measured in triplicate with a transwell system (cat. no.3421, 24-well plates; 5.0 µm pore size; Corning Costar, NY). RPMI 1640 with 1% FCS alone, with varying doses of M-CSF or IL34 (cat. no. 200-34, PeproTech) added to the lower chamber. Wells with medium alone were used as a control for spontaneous migration. A total of 2.5 × 10 5 cells in 200 µl was added to the upper chamber and incubated at 37°C for 4 h. In some migration experiments, cells were pretreated with GW2580 (1uM, cat. no. SML1047, Sigma-Aldrich) or with BLZ945 (0.5 M, Selleckchem) for 60 min at 37°C. For migration of BDCA1 + LC, 1.5 × 10 5 cells in 200 µl were added to the upper chamber of a transwell system (5.0 µm pore size; Corning Costar) and harvested after 6 h of migration towards M-CSF (50 ng/ml). Mo-LCs and BDCA1 + LCs were not enriched for CD207. Cells migrated into the lower chamber were harvested, concentrated to a volume of 300 µl of PBS, and counted by flow cytometry. Events were acquired for a fixed time of 60 s. The counts fell within a linear range of the control titration curves obtained by testing increasing cell concentrations. Values are given as the mean number of migrated cells ± SEM.

Lonardi S., Scutera S., Licini S., Lorenzi L., Cesinaro A.M., Gatta L.B., Castagnoli C., Bollero D., Sparti R., Tomaselli M., Medicina D., Calzetti F., Cassatella M.A., Facchetti F., Musso T, & Vermi W. (2020). CSF1R Is Required for Differentiation and Migration of Langerhans Cells and Langerhans Cell Histiocytosis. Cancer immunology research, 8(6).

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Hypoxia-induced Cell Culture Assay

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U87 and U251 cells were cultured in DMEM (10% FBS), and THP-1 cells were cultured in RPMI-1640 (10% FBS). All cells were maintained in 20% O₂ at 37°C in a 5% CO₂ humidified chamber. Hypoxic conditions were induced by incubating the cells under 1% O₂, 5% CO₂, and 94% N₂ at 37°C. PD153035 (BioVision), LY294002 (Beyotime Biotechnology), BLZ945 (selleck), TGF-α (PeproTech) and Recombinant POSTN (Biovision) were purchased.

Guo X., Xue H., Shao Q., Wang J., Guo X., Chen X., Zhang J., Xu S., Li T., Zhang P., Gao X., Qiu W., Liu Q, & Li G. (2016). Hypoxia promotes glioma-associated macrophage infiltration via periostin and subsequent M2 polarization by upregulating TGF-beta and M-CSFR. Oncotarget, 7(49), 80521-80542.

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Innate Immune Signaling Modulation

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High molecular weight (HMW) Poly(I:C), LPS, 2′3′-cGAMP, HMW Poly(I:C) complexed with LyoVec transfection reagent, and Bx795 (all Invivogen); IKK16 and Amlexanox (Tocris Bioscience), BLZ945 (Selleckchem), IFNα2 and mouse IFNα3 (PBL Assay Science), were reconstituted per manufacturer’s instructions; concentrations are noted in figure legends. SiRNA against MDA5 or All-Stars negative control siRNA (Qiagen) were complexed with lipofectamine 2000 (Thermo-Fisher) following manufacturer’s instructions, delivering 100pmol of siRNA per 35 mm dish, and changing media after 6 h. Infections/treatments of siRNA-treated MDMs occurred 36 h after transfection.

Brown M.C., Mosaheb M.M., Mohme M., McKay Z.P., Holl E.K., Kastan J.P., Yang Y., Beasley G.M., Hwang E.S., Ashley D.M., Bigner D.D., Nair S.K, & Gromeier M. (2021). Viral infection of cells within the tumor microenvironment mediates antitumor immunotherapy via selective TBK1-IRF3 signaling. Nature Communications, 12, 1858.

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EGFR Mutant NSCLC Cell Line Protocol

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EGFR mutant (del E746_A750) NSCLC cell lines PC9 were obtained from Dr. Nishio Kazuto (Kindai University, Osaka, Japan) in 2005. PC9 were grown in RPMI-1640 (Gibco) with 10% FBS (Gemini) and 1% penicillin/streptomycin (Gibco). The murine NIH/3T3 cells were purchased from the ATCC in 2010 and were maintained in DMEM (Gibco) with 10% FCS (Sigma-Aldrich) and 1% penicillin/streptomycin. PC9 cells were authenticated in 2017 using the Promega GenePrint 10 System at the RTSF Research Technology Support Facility in the
Genomic Core Laboratory, Michigan State. Murine NIH/3T3 cells were not authenticated. All cell lines were tested negative for Mycoplasma using the Mycoplasma Plus PCR Primer Set (Agilent). Osmertinib, sunitinib, gilteritinib, axitinib, BLZ945, nintedanib, cabozantinib, OTX015, RAF709, trametinib, and SCH772984 were purchased from Selleck Chemicals. Loxo292 was purchased from ProbeChem (Shanghai, China).

Cooper A.J., Kobayashi Y., Kim D., Clifford S.E., Kravets S., Dahlberg S.E., Chambers E.S., Li J., Rangachari D., Nguyen T., Costa D.B., Rabin M.S., Wagle N., Sholl L.M., Jänne P.A, & Oxnard G.R. (2020). Identification of a RAS-activating TMEM87A-RASGRF1 fusion in an exceptional responder to sunitinib with non-small cell lung cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(15), 4072-4079.

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