Rottlerin

Manufactured by Merck Group

Sourced in United States, Germany, China

Rottlerin is a laboratory reagent produced by Merck Group. It is a small molecule compound commonly used in scientific research. Rottlerin's core function is to serve as a selective inhibitor of the protein kinase C delta (PKC-δ) enzyme.

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Lab products found in correlation

50 protocols using rottlerin

Colitis Induction and Rottlerin Treatment in Mice

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WT mice (male, ten weeks old) were randomly allocated to three groups (the normal group, the vehicle group, which received only DSS (dextran sulfate sodium), and the treatment group, which received DSS + rottlerin) with 10 mice per group. DSS was dissolved in drinking water, the colitis was induced by DSS (MP Biomedicals, Cat #: 160110) for 2 (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
circulations. For the first circulation, DSS was administered for 4 days, and then normal drinking water was given for another 10 days; based on previous reports [19] , the oral administration of rottlerin (100 mg/kg, Sigma Aldrich, Cat #: R5648) began after the administration of DSS in the first circulation [20] . rottlerin was dissolved as a suspension in 0.9% NaCl (Sigma Aldrich, Cat #: 793566) and orally administered once daily until the end of the second circulation (the steps of the second circulation were the same as those of the first).
After 4 weeks, the mice were humanely sacrificed and the entire colon of each mouse was collected.

Song X., Zuo L., Wang L., Zhu Z., Tao J., Jiang Y., Wu X., Wang Z., Nian J., Xiang P., Zhang Q., Zhao H., Liang Y., Li J., & Hu J. (2020). Rottlerin ameliorates DSS-induced colitis by improving intestinal barrier function via activation of the Epac-2/Rap-1 signaling pathway.

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Culturing Normal Human Astrocytes and U-87 Astrocytoma Cells

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Normal human astrocytes (NHA) isolated from the cerebrums of 5-month-old human fetuses were purchased from Cambrex (CC-2565, Walkersville, MD, USA), and cultured according to the manufacturer’s protocol. The astrocytoma human cell line U-87 was routinely grown in cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Rockville, MD, USA) containing 10% heat-inactivated fetal calf serum, 1% penicillin/streptomycin, and 2 mM L-glutamine (ICN Pharmaceuticals, CA, USA) at 37 °C in a humidified atmosphere of 5% CO₂.
Bryostatin-1, prostratin, GF109203X, and rottlerin were purchased from Sigma (St. Louis, MO, USA). Pyrrolidine dithiocarbamate (PDTC), and BAY11-7082 were obtained from Santa Cruz Biotechnology (Santa Cruz CA, USA).

Díaz L., Martínez-Bonet M., Sánchez J., Fernández-Pineda A., Jiménez J.L., Muñoz E., Moreno S., Álvarez S, & Muñoz-Fernández M.Á. (2015). Bryostatin activates HIV-1 latent expression in human astrocytes through a PKC and NF-ĸB-dependent mechanism. Scientific Reports, 5, 12442.

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Cellular Imaging Reagents Protocol

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5-(N-Ethyl-N-isopropyl) amiloride (EIPA), blebbistatin, casin, chlorpromazine, cytochalasin D, dynasore, filipin complex, gefitinib (Iressa), genistein, IPA-3, nocodazole, NSC23766, rottlerin, salirasib, wortmannin, Y11, Fluoromount^TM, polyethylene glycol 8000 and Sepharose® 6B were from Sigma-Aldrich (St. Louis, MO, USA). Alexa Fluor® 633-conjugated transferrin, Lysine fixable Dextran-Texas Red®, 70,000 MW, Alexa Fluor® 594-conjugated cholera toxin subunit B and rhodamine phalloidin were from Molecular Probes, Thermo Fisher Scientific (Waltham, MA USA).

Levican J., Miranda-Cárdenas C., Soto-Rifo R., Aguayo F., Gaggero A, & León O. (2017). Infectious pancreatic necrosis virus enters CHSE-214 cells via macropinocytosis. Scientific Reports, 7, 3068.

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Subcellular Localization of Au-shRNA Nanocarriers

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HepG2 cells were plated (1 × 10⁶ cells per well) on a laser confocal culture dish, and the cells were first pretreated with various endocytosis inhibitors for about 1 h, then, gold salt and shRNA were added sequentially and incubated with the cells for 6 h. The concentrations of the inhibitors was as follows: 37 mg/mL methyl-β-cyclodextrin, 10 μg/mL chlorpromazine, 10 mg/mL rottlerin, 200 μg/mL genistein, and 5 μg/mL filipin III (Sigma-Aldrich, MO, USA). After 6 h of co-incubation, the HepG2 cells were washed three times with PBS and fixed with 4% paraformaldehyde for 30 min. The cell nucleus was stained with DAPI (Beyotime, Shanghai, China). Finally, a confocal microscope was used to image the sample with 488 nm (Leica, Wetzlar, Germany).
To observe the subcellular localization of Au–shRNA NCs, gold salt and the shRNA were co-incubated with HepG2 cells at 37 °C for 12 h. The specific experimental process was the same as that above-mentioned. Endosomes and lysosomes were labeled by Lysotracker Red for 30 min and washed with PBS (three times), followed by nuclei staining with DAPI for 3 min. The images were obtained by confocal microscopy (Leica, Wetzlar, Germany).

Cai W., Yin L., Jiang H., Weizmann Y, & Wang X. (2021). Intelligent Bio-Responsive Fluorescent Au–shRNA Complexes for Regulated Autophagy and Effective Cancer Bioimaging and Therapeutics. Biosensors, 11(11), 425.

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Aptamer-based Detection of SGIV Infection

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The aptamer Q5 was generated against SGIV-infected cells with Cell-SELEX in our previous study (Li et al., 2015 (link)). Aptamer Q5 was synthesized by Life Technologies, and the 5′ end of aptamer Q5 was labeled fluorescently with Cy5 (Cy5-Q5). Chlorpromazine (CPZ), methyl-β-cyclodextrin (MβCD), nystatin, genistein, dynasore, (N-ethyl-N-isopropyl)-amiloride (EIPA), ML-7, NSC23766, rottlerin, IPA-3, chloroquine (CQ), ammonia chloride (NH₄Cl), cytochalasin D (cytoD), and nocodazole were purchased from Sigma-Aldrich (St. Louis, MO, United States). All reagents were stored and dissolved to specific concentrations according to the manufacturer’s instructions.

Yu Q., Liu M., Wu S., Wei X., Xiao H., Yi Y., Cheng H., Wang S., Zhang Q., Qin Q, & Li P. (2020). Specific Aptamer-Based Probe for Analyzing Biomarker MCP Entry Into Singapore Grouper Iridovirus-Infected Host Cells via Clathrin-Mediated Endocytosis. Frontiers in Microbiology, 11, 1206.

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PaCa Cell Line Rottlerin Treatment

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The human PaCa cell lines employed were obtained from American Type Culture Collection (Manassas, VA, USA). PANC-1 and MIAPaCa-2 cells were cultured in DMEM/F12 supplemented with 10% FBS. All media contain penicillin and streptomycin (100 units/ml). Cells were maintained at 37°C in a humidified atmosphere containing 5% CO₂/95% air, and were used between passages 4 and 15. Rottlerin was purchased from (Sigma-Aldrich, St. Louis, MO, USA), dissolved in DMSO and directly added to the cell cultures at indicated concentrations (μM). Control cells were treated with DMSO alone.

Ashour A.A., Gurbuz N., Alpay S.N., Abdel-Aziz A.A., Mansour A.M., Huo L, & Ozpolat B. (2014). Elongation factor-2 kinase regulates TG2/β1 integrin/Src/uPAR pathway and epithelial–mesenchymal transition mediating pancreatic cancer cells invasion. Journal of Cellular and Molecular Medicine, 18(11), 2235-2251.

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Macrophage Differentiation Protocol

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PMA was purchased from Sigma (#P8139). Recombinant human IFNγ was purchased from
BD Biosciences (#554617). Lipopolysaccharide Escherichia coliO111:B4 was purchased from Sigma (#L2630). Rottlerin was purchased from Sigma
(#R5648). Recombinant human granulocyte-macrophage colony-stimulating factor
(GM-CSF) was purchased from Sanofi (Leukine sargramostim). Recombinant human
macrophage colony-stimulating factor 1 (M-CSF) was purchased from GenScript
(#Z02001). Recombinant human interleukin-4 (IL-4) was produced at the Ludwig
Cancer Research (Brussels, Belgium). Recombinant human IL-10 was purchased from
R&D (#1064-IL). Ribomunyl was purchased from Pierre Fabre (France).
Prostaglandin E2 (PGE2) was purchased from Sigma. Recombinant human tumor
necrosis factor alpha (TNFα) was purchased from PeproTech (#300-01A).
Recombinant human transforming growth factor beta (TGFβ) was purchased from
R&D (#240-B).

Hoffmann D., Pilotte L., Stroobant V, & Van den Eynde B.J. (2019). Induction of tryptophan 2,3-dioxygenase expression in human monocytic leukemia/lymphoma cell lines THP-1 and U937. International Journal of Tryptophan Research : IJTR, 12, 1178646919891736.

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Antibodies for Western Blot Analysis

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Rottlerin (Sigma-Aldrich; Merck Millipore) was dissolved in dimethyl sulfoxide (DMSO) as a stock solution of 20 mM and stored at −20°C. Human microtubule-associated protein 1 light chain 3 (LC3; cat. no. ABC432; working dilution, 1:1,000) antibody was purchased from Sigma-Aldrich (Merck Millipore). Human caspase-3 (cat. no. ab13585; dilution, 1:1,000) and human poly (ADP-ribose) polymerase (PARP; cat. no. ab75607; dilution, 1:400) antibodies were purchased from Abcam (Cambridge, MA, USA). The human anti-β-actin antibody (cat. no. sc-130065; dilution, 1:1,000) was obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Horseradish peroxidase-conjugated donkey anti-rabbit (cat. no. NA934; dilution, 1:1,000) and anti-mouse (cat. no. N1034; dilution, 1:10,000) IgG secondary antibodies were obtained from GE Healthcare Life Sciences (Uppsala, Sweden).

Qi P., He Z., Zhang L., Fan Y, & Wang Z. (2016). Rottlerin-induced autophagy leads to apoptosis in bladder cancer cells. Oncology Letters, 12(6), 4577-4583.

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Investigating Topotecan-Induced Apoptosis Pathways

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NSC, and topotecan were kindly provided by the Drug Synthesis and Chemistry Branch, Developmental Therapeutic Program, National Cancer Institute (Bethesda, MD, USA). Tissue culture medium, rottlerin, protease inhibitors, propidium iodide (PI), and anti-β-actin primary antibody were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS), L-glutamine, penicillin and streptomycin were from Gemini Bio-Products (Calabasas, CA, USA). Z-VAD-fluoromethylketone (FMK), RNase A, protease K, and M-MLV reverse transcriptase were from Promega (Madison WI, USA). Antibodies against cyclin A, and PKCδ were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The cytochrome c antibody (clone 7H8.2C12) was obtained from BD Pharmingen (San Diego, CA, USA). The SYBR-Green Real-Time PCR master mix was from Life Technologies (Grand Island, NY, USA). TriPure Isolation reagent was from Roche Applied Science (Mannheim, Germany).

Wang X., Tan C., Wang G., Cai J.J., Wang L.P., Imperato-McGinley J, & Zhu Y.S. (2017). Dual action of NSC606985 on cell growth and apoptosis mediated through PKCδ in prostatic cancer cells. International Journal of Oncology, 51(5), 1601-1610.

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Preparation of Bioactive Compound Solutions

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GO6976 (GO), LY333531 (LY), Rottlerin (Rott), rosmarinic acid, and apigenin were purchased from Sigma Chemical Co. (Yongin, Korea). Salvianolic acid B was purchased from Santa Cruz (Santa Cruz, CA, USA). Stock solutions of GO, LY, Rott, and apigenin were dissolved in dimethyl sulfoxide (DMSO). Salvianolic acid B and rosmarinic acid were dissolved in distilled water. The final concentration of DMSO was less than 0.1%.

Kwon O.J., Oh H.C., Lee Y.J., Kim H.Y., Tan R., Kang D.G, & Lee H.S. (2015). Sibjotang Increases Atrial Natriuretic Peptide Secretion in Beating Rabbit Atria. Evidence-based Complementary and Alternative Medicine : eCAM, 2015, 268643.

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