Human skin fibroblasts (HSF), keratinocytes (HaCaT), and human umbilical vein endothelial cells (HUVECs), purchased from ATCC (Manassas, VA, USA), were cultured in DMEM (Gibco, Carlsbad, CA, USA) containing 10% FBS (Gibco) at 37°C with 5% CO2. For β‐catenin inhibition, HSF cells and HUVECs were treated with a β‐catenin inhibitor (JW 55, 5 μM) (Medchemexpress, Shanghai, China) for 24 h. For CYP1B1 inhibition, HSF cells and HUVECs were treated with a CYP1B1 inhibitor (CYP1B1‐IN‐1, 10 μM; Medchemexpress) for 24 h.
Huvecs
Manufactured by MedChemExpress
Sourced in United States, China
HUVECs (Human Umbilical Vein Endothelial Cells) are primary endothelial cells derived from the human umbilical vein. They are a widely used in vitro cell model for studying endothelial cell biology and function.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Huvecs, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Hy 100941, by MedChemExpress (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Huvecs, by BNCC (1 mentions)
Rosuvastatin, by MedChemExpress (1 mentions)
Mitomycin c, by MedChemExpress (1 mentions)
Huvecs, by GenePharma (1 mentions)
Dmi8 microscope, by Leica (1 mentions)
Huvecs, by PromoCell (1 mentions)
Jw 55, by MedChemExpress (1 mentions)
Jsh 23, by MedChemExpress (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Sunitinib, by MedChemExpress (1 mentions)
6 protocols using huvecs
1
Inhibition of β-catenin and CYP1B1 in Skin Cells
2
Sal B Protects HUVECs from HG-Induced Damage
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Human umbilical vein endothelial cells (HUVECs; ATCC, USA) were grown in Dulbecco’s modified Eagle’s medium (Gibco, USA) plus 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 U/ml streptomycin. HUVECs were grown in an environment of 5% CO2 at 37°C. Diabetes cell model was constructed by 30 mM high-glucose (HG)-induced HUVECs. 0.1% DMSO was utilized for improving the solubility and bioavailability of Sal B. HUVECs were treated with 0, 5, 10, 30, 50, 80 and 100 μM Sal B for 48 h in HG-pre-treated HUVECs. Furthermore, HUVECs were treated with 50 nM oxidative phosphorylation uncoupler carbonyl cyanide m-chlorophenyl hydrazone (CCCP; HY-100941; Med Chem Express, USA) that was dissolved by 0.1% DMSO.
3
Rosuvastatin Pretreatment Attenuates H2O2-induced Oxidative Stress in HUVECs
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HUVECs were obtained from the BeNa Culture Collection (Beijing, China). The cells were cultured in Dulbecco’s modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (Gibco, United States), streptomycin (100 μg/ml), and penicillin (100 IU/ml) in a humidified atmosphere containing 5% CO2/95% air at 37°C. The culture medium was replaced every 2 days.
In the experiment, HUVECs were pretreated with rosuvastatin (MedChemExpress, United States; 2.5, 5, and 10 μM) for 2 h. The concentrations were determined prior to the experiment with a drug concentration gradient (Wang et al., 2010 (link)). rosuvastatin was dissolved at a certain concentration in dimethyl sulfoxide (DMSO) (Piconi et al., 2008 (link)); the final concentration of DMSO was always lower than 0.01%, which had been shown to have no effect on cell viability (Gao et al., 2008 (link)). The cells were then exposed to H2O2 (750 μM) for 24 h (Chen et al., 2014 (link)). The final concentrations of H2O2 were determined by the experiment with a H2O2 concentration gradient.
In the experiment, HUVECs were pretreated with rosuvastatin (MedChemExpress, United States; 2.5, 5, and 10 μM) for 2 h. The concentrations were determined prior to the experiment with a drug concentration gradient (Wang et al., 2010 (link)). rosuvastatin was dissolved at a certain concentration in dimethyl sulfoxide (DMSO) (Piconi et al., 2008 (link)); the final concentration of DMSO was always lower than 0.01%, which had been shown to have no effect on cell viability (Gao et al., 2008 (link)). The cells were then exposed to H2O2 (750 μM) for 24 h (Chen et al., 2014 (link)). The final concentrations of H2O2 were determined by the experiment with a H2O2 concentration gradient.
4
Knockdown and Overexpression of PTBP1 in HUVECs
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Human umbilical vein cells (HUVECs, PromoCell, C-12208) were cultured in endothelial cell growth medium (PromoCell, C-22010). To knockdown the expression of PTBP1, HUVECs were infected with lentivirus-shRNA-PTBP1 (LV-sh-PTBP1) (Gene Pharma, Inc.) at a multiplicity of infection (MOI) of 50 at 37 °C for 24 h, and then cultured with fresh medium for another 48 h at 37 °C in a 5% CO2 incubator before subsequent experiments. The cells infected with control lentivirus-shRNA (Gene Pharma, Inc.; MOI, 50) were used as a negative control. For the migration assay, HUVECs were incubated with 10 μg/mL mitomycin C (MedChemExpress) at 37 °C in a 5% CO2 atmosphere for 2 h50 (link). The LV-sh-PTBP1 sequences were as follows: 5’-CGGCACAGTGTTGAAGATCAT-3’. For the overexpression of ARRB1-L or ARRB1-S, HUVECs were infected with ARRB1-L overexpression lentivirus or ARRB1-S overexpression lentivirus. At 72 h after infection, the growth area was scratched. The cells were then photographed in five different fields at 0 h and 20 h after scratching using a Leica DMi8 microscope, respectively. Cell migration ability was calculated as a percentage of the wound area difference between 0 h and 20 h to the wound area at 0 h by ImageJ software.
5
Investigating HDAC1 Inhibition and NF-κB Signaling in Atherosclerosis
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HUVECs (ATCC CRL-1730) were purchased from American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM (Thermo Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (FBS; Thermo Fisher Scientific). The cells were cultured at the controlled temperature of 37°C in a 5% CO2 atmosphere. The cells in the logarithmic growth phase were extracted and seeded into a 6-well plate (3 × 105 cells/well). When the cell confluence reached 50%, the cells were transfected using Lipofectamine 2000 (11668019; Invitrogen, Carlsbad, CA, USA). Subsequent to 24 h of transfection, the cells were further treated with 50 μg/mL ox-LDL (Beijing Xiesheng Bio-Technology, Beijing, China) for an additional 24 h.
To verify the effect of NF-κB signaling on atherosclerosis, HDAC1 was inhibited in HUVECs, and IKB JSH-23 (HY-13982; MedChemExpress, NJ, USA; 20 μm) was added to the cells.15 (link) The experiment was divided into three groups: sh-NC + DMSO, sh-HDAC1 + DMSO, and sh-HDAC1 + JSH-23. Subsequent to the transfection, the expressions of HDAC1, p65, and p-p65 were detected using the western blot analysis, the cell activity was detected by the CCK-8, HUVEC survival was detected by clone formation, and the apoptosis was detected using flow cytometry.
To verify the effect of NF-κB signaling on atherosclerosis, HDAC1 was inhibited in HUVECs, and IKB JSH-23 (HY-13982; MedChemExpress, NJ, USA; 20 μm) was added to the cells.15 (link) The experiment was divided into three groups: sh-NC + DMSO, sh-HDAC1 + DMSO, and sh-HDAC1 + JSH-23. Subsequent to the transfection, the expressions of HDAC1, p65, and p-p65 were detected using the western blot analysis, the cell activity was detected by the CCK-8, HUVEC survival was detected by clone formation, and the apoptosis was detected using flow cytometry.
6
Culturing HUVECs with Lyc, Sunitinib, and PDGF-AA
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Human Umbilical Vein Endothelial Cells (HUVECs) were purchased from Cell Bank of Representative Culture Preservation Committee of Chinese Academy of Sciences (Shanghai, China). HUVECs were cultured in HUV-EC-C medium and kept in an incubator at 37 °C with 5% CO2.
Lyc was purchased from MedChemExpress (HY-N0288) and prepared as described before [27 (link)]. Sunitinib and PDGF-AA were also purched from MedChemExpress (HY-10255A; HY-P70598).
Lyc was purchased from MedChemExpress (HY-N0288) and prepared as described before [27 (link)]. Sunitinib and PDGF-AA were also purched from MedChemExpress (HY-10255A; HY-P70598).
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