Human SK-Hep-1 cells derived from the ascitic fluid of liver adenocarcinoma were purchased from the American Type Culture Collection. Human Hep-J5, Huh-7, and Mahlavu cells were obtained from Dr. S-N Wang (KMU, Taiwan). Human Hep-G2 cells and Hep-3B cells were purchased from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). The cells were cultured in the Dulbecco's modified Eagle medium (DMEM) (Gibco, Grand Island, NY, USA) containing 10% heat-inactivated fetal bovine serum (FBS, Gibco, Grand Island, NY, USA) and 100 U/mL penicillin/streptomycin at 37°C in an incubator with 5% CO2. The cells were transfected with 50 nM of siRNA targeting HDAC6 (sc-35544, Santa Cruz Biotechnology Inc., CA, USA), HDAC6-Flag (Addgene plasmid number #13823), dominant-negative HDAC6.DC-FLAG (Addgene plasmid number #30483), or HDAC6-Nuc [12] (link) using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions.
Hep3b cells
Manufactured by RIKEN BioResource Center
Hep3B cells are a human hepatocellular carcinoma cell line derived from the liver tissue of a patient. They are widely used in biomedical research as a model system for studying liver function and disease.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Hdac6 flag, by Addgene (1 mentions)
Transit lt1 transfection reagent, by Mirus Bio (1 mentions)
Mcf 7, by RIKEN BioResource Center (1 mentions)
Mda mb 231, by RIKEN BioResource Center (1 mentions)
Hek293 cells, by RIKEN BioResource Center (1 mentions)
Trizma base solution, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Cisplatin, by Merck Group (1 mentions)
4 protocols using hep3b cells
1
Transfection of Liver Cancer Cell Lines
2
Evaluating miR-4669 Impact on Hep3B Cells
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Hep3B cells were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan). To explore the impact of miR-4669 overexpression on cell viability and migration, Hep3B cells were transfected with a miR-4669 mimic (10, 20, or 50 nM) or a negative control mimic (Qiagen, Hilden, Germany) according to a previous study [16 (link)]. After transfection for 24 h, cell viability was evaluated using a Cell Counting Kit-8 (CCK-8) assay (MilliporeSigma, Burlington, MA, USA). Migration assays were performed according to a previous study [16 (link)], and migrated cells at the bottom of the Transwell membrane (8 μm pore size, MilliporeSigma) were stained with 0.2% crystal violet for cell counting.
3
Exosome Uptake and Effects in Cultured Cancer Cells
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Human hepatoma Huh-7, Hep3B cells, human embryonic kidney Hek293 cells, breast carcinoma MCF-7, and MDA-MB-231 cells were all purchased from Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan) with authentication. They were maintained in DMEM medium supplemented with 10% (v/v) fetal bovine serum (FBS) at 37° C under 5% CO2 and 100 % humidity. Plasmid transfection was done using transit-LT1 transfection reagent (Mirus Bio LLC., Medison, WI, USA) according to the manufacturer's instruction. Transfected cells were selected and enriched with culture medium containing G418, hygromycin B, or puromycin. For the treatment of exosome at MCF-7 cells, 2 × 105 cells were plated overnight and incubated at serum-free medium for 4 hr. Exosome solution with desired protein equivalent was added and pipetted gently into the medium and kept at incubator for additional 24 hr. The cells were then harvested for PCR, or western blot.
4
Cytotoxicity Assay of Cisplatin in HCC Cells
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The two HCC cell lines used in the present study, Hep3B cells purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan) and Mahlavu cells provided by Professor K.H. Lin (Chang Gung University, Taiwan), were grown in Dulbecco's modified Eagle's medium (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum (Biological Industries, Cromwell, CT, USA) in 5% CO2, in a humidified incubator (37°C). The cells were treated with cisplatin (Sigma-Aldrich; Merck Millipore, Darmstadt, Germany) at various concentrations for 24 or 72 h, and cell viability was measured using a sulforhodamine B (SRB) assay, as previously described (12 (link)). Briefly, cells were fixed using trichloroacetic acid and then incubated with SRB (Sigma-Aldrich; Merck Millipore) upon being washed and air-dried. The precipitate was dissolved in Trizma® base solution (Sigma-Aldrich; Merck Millipore). The absorbance was measured at 540 nm by using a microplate reader.
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