Doxycycline hyclate

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Doxycycline hyclate is a tetracycline antibiotic compound. It is a crystalline powder used as a reagent in laboratory applications.

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Doxycycline (23 citations)

4 protocols using doxycycline hyclate

Culturing tau-expressing SH-SY5Y neurons

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The neuronal-like cell line SH-SY5Y-eGFP-tau-P301L (SY5Y-tau) (26 (link)) was a gift from Yung-Feng Liao at the Institute of Cellular and Organismic Biology, Academia Sinica. SY5Y-tau cells were grown in DMEM (HyClone) supplemented with 10% heat-inactivated FBS, 5 μg/mL blasticidin (Gibco) and 200 μg/mL hygromycin (Thermo Fisher Scientific) at 37 °C in a humidified 5% CO₂-containing atmosphere. All SY5Y-tau cells were induced to express tau for subsequent experiments with 1 μg/mL doxycycline hyclate (Santa Cruz).

Siew J.J., Chen H.M., Chiu F.L., Lee C.W., Chang Y.M., Chen H.L., Nguyen T.N., Liao H.T., Liu M., Hagar H.T., Sun Y.C., Lai H.L., Kuo M.H., Blum D., Buée L., Jin L.W., Chen S.Y., Ko T.M., Huang J.R., Kuo H.C., Liu F.T, & Chern Y. (2024). Galectin-3 aggravates microglial activation and tau transmission in tauopathy. The Journal of Clinical Investigation, 134(2), e165523.

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Compound Preparation for Cell Experiments

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A total of 100 mM of Ivabradine (Sigma), 100 mg/mL of puromycin (Sigma), 50 mM of sodium phenylbutyrate (4‐PBA; Santa Cruz Biotechnology) and 50 mg/mL of doxycycline hyclate (Santa Cruz Biotechnology) were dissolved in double‐distilled water. A total of 50 μM of Paclitaxel (Selleckchem) was dissolved in DMSO. Water and/or saline were used as solvent control in in vitro and in vivo experiments. Also, 10 mM of Inositol 1,4,5‐Trisphosphate (407137; Sigma) in water was used. A sum of 1 mM of Fura‐2‐AM (Invitrogen) stock solution in DMSO was prepared and 10 mg/mL of Hoechst33342 was purchased from ThermoFisher.

Mok K., Tsoi H., Man E.P., Leung M., Chau K.M., Wong L., Chan W., Chan S., Luk M., Chan J.Y., Leung J.K., Chan Y.H., Batalha S., Lau V., Siu D.C., Lee T.K., Gong C, & Khoo U. (2021). Repurposing hyperpolarization‐activated cyclic nucleotide‐gated channels as a novel therapy for breast cancer. Clinical and Translational Medicine, 11(11), e578.

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Breast cell line cultivation and compound treatment

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All the cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and were confirmed by short tandem repeat (STR) identification. Two breast epithelial cell lines, MCF10A and 184A1, were cultured in Dulbecco’s modified Eagle’s medium (DMEM)-F12 medium, supplemented with 5% fetal bovine serum (FBS), 10 μg/mL insulin, 20 ng/mL EGF, 100 ng/mL cholera toxin, 0.5 μM calcium chloride, and 500 ng/mL hydrocortisone. All other cell lines were cultured in DMEM-F12 medium containing 10% FBS in a 37°C incubator supplemented with 5% CO₂. Antibiotics (penicillin and streptomycin) were added to the medium according to the manufacturer’s instructions.
Compounds including SP600125, trametinib, MK2206, saracatinib, CHX, MG132, erlotinib, gefitinib, afatinib, dacomitinib, rociletinib, and osimertinib were purchased from Selleck Chemicals. Doxycycline hyclate was obtained from Santa Cruz Biotechnology.

Li Y., Tan Y., Wen L., Xing Z., Wang C., Zhang L., Wu K., Sun H., Li Y., Lei Q, & Wu S. (2021). Overexpression of BIRC6 driven by EGF-JNK-HECTD1 signaling is a potential therapeutic target for triple-negative breast cancer. Molecular Therapy. Nucleic Acids, 26, 798-812.

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Inducible Stable Cell Lines for TPX2 Study

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Stable transgenic hTERT RPE-1 cell lines were generated by transfection of a plasmid encoding the piggyBac transposase together with inducible vectors for expression of VENUS alone (epB-Bsd-TT-VENUS), FLAG-TPX2 full length/VENUS (epB-Puro-TT-FLAG-TPX2 and epB-Bsd-TT-VENUS), FLAG-Δ43TPX2/VENUS (epB-Puro-TT-FLAG-Δ43TPX2 and epB-Bsd-TT-VENUS) or FLAG-Δ43TPX2 alone (epB-Puro-TT-FLAG-Δ43TPX2 and epB-Bsd-TT). Transfection was performed using Lipofectamine LTX (Invitrogen, Carlsbad, CA, USA). Then, 48 h after transfection, selection with blasticidin-S hydrochloride and puromycin (both 9 μg/mL; Sigma-Aldrich, St Louis, MO, USA) was applied. Resistant cells were propagated as a pool, and expression of exogenous proteins after administration of 1 μg/mL doxycycline hyclate (dox, tetracycline analogue; Santa Cruz Biotechnology, Dallas, TX, USA) was verified.

Naso F.D., Sterbini V., Crecca E., Asteriti I.A., Russo A.D., Giubettini M., Cundari E., Lindon C., Rosa A, & Guarguaglini G. (2020). Excess TPX2 Interferes with Microtubule Disassembly and Nuclei Reformation at Mitotic Exit. Cells, 9(2), 374.

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