Human umbilical vein endothelial cells (HUVECs) were purchased from Zhejiang Ruyao Biotechnology Co., Ltd. HUVECs were cultured in EGM-2 medium (cat no. CC-3162, Lonza) for endothelial cells. Subculturing was performed in a 37 °C incubator containing 5% CO2. Cell grouping (cell group 1) was performed as follows: 1. HUVEC group, experimental control, without any treatment; 2. HG + ox-LDL group, supplied 33.3 mM d -glucose (You et al. 2019 (link)) + 100 μg/mL ox-LDL (Zhang et al. 2019 (link)) [ox-LDL was purchased from Yeasen Biotechnology (Shanghai) Co., Ltd.; cat no. 20605ES05] for 48 h; 3. HSYA group, HUVECs were treated with 25 μM HSYA for 48 h; 4. HG + ox-LDL + HSYA group, 33.3 mM d -glucose + 100 μg/mL ox-LDL was administered, and then the cells were simultaneously treated with 25 μM HSYA for 48 h. After the experiment, the GSH-Px, MDA, ROS, and iron ion contents in the cell lysate were detected. The expression levels of the ferroptosis-related proteins SLC7A11, GPX4, and ACSL4 were also detected using western blotting.
Ox ldl
Manufactured by Yeasen
Sourced in China
Ox-LDL is a laboratory equipment used for the measurement of oxidized low-density lipoprotein (Ox-LDL) levels. Ox-LDL is a biomarker associated with the development of atherosclerosis and cardiovascular disease.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
F12k medium, by Procell (2 mentions)
Rnaimax, by Thermo Fisher Scientific (2 mentions)
Phorbol myristate acetate (pma), by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Yeasen (1 mentions)
Ha vsmc, by American Type Culture Collection (1 mentions)
Ml385, by Selleck Chemicals (1 mentions)
Huvecs, by Procell (1 mentions)
Ham s f12k, by Procell (1 mentions)
Heparin, by Procell (1 mentions)
Oil red o staining kit, by Beyotime (1 mentions)
Calcein am, by BioLegend (1 mentions)
Transwell apparatus, by Corning (1 mentions)
Tcs sp8 x, by Leica camera (1 mentions)
Foetal bovine serum (fbs), by Corning (1 mentions)
Human umbilical vein endothelial cells huvecs, by ScienCell (1 mentions)
Penicillin streptomycin, by Corning (1 mentions)
U937 cells, by American Type Culture Collection (1 mentions)
Ox ldl, by Merck Group (1 mentions)
22 protocols using ox ldl
1
HSYA Mitigates Endothelial Cell Damage
2
Ox-LDL Induced VSMC Cytotoxicity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VSMCs were diluted in F12K medium (Procell, Wuhan, China) and grown in 96-well plates for 16h. Then, cells were incubated with various concentrations (0, 25, 50, 75 and 100g/mL) of ox-LDL (Yeasen, Shanghai, China) for 24h or 50g/mL ox-LDL for different time (0, 12, 24, 36 and 48h). After that, MTT reagent (Solarbio, Beijing, China) was incubated with cells for 4h. Cell supernatant was discarded and dimethyl sulfoxide (Sigma, St. Louis, MO, USA) was used to dissolve formazan. The cell viability was determined by assessing the output of wavelength at 490 nm with a Varioskan LUX Multimode microplate reader (Thermo Fisher, Waltham, MA, USA).
3
Ox-LDL-induced HUVEC Transcription Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cultured HUVECs were stimulated using 100 mg/L Ox-LDL (Shanghai Yeasen Biotechnology Co. Ltd. Shanghai, China) in endothelial basal medium for 24 h, and Ox-LDL-induced HUVECs were treated with 0, 4, 20, 100, 200, and 300 μM AF for 24 h, respectively. Vector (control) and IRAK1-overexpressed plasmids were purchased from Origene (Rockville, MD, USA). Ox-LDL-induced HUVECs were evenly placed into a 6-well plate and cultured for 12 h at 37°C. Then, the cells were transfected with the vector and IRAK1-overexpressing plasmid with Lipofectamine 3000 (Invitrogen, Waltham, MA, USA) on the basis of the instructions.
4
Atherosclerosis Model via Ox-LDL Treatment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human VSMCs from the National Infrastructure of Cell Line Resource (cat. no. 4201HUM-CCTCC00632) were cultured in DMEM (Thermo Fisher Scientific, Inc.) containing 10% FBS (Thermo Fisher Scientific, Inc.) and incubated at 37˚C with 5% CO2. To construct the atherosclerosis model in vitro, VSMCs were treated with ox-LDL (Shanghai Yeasen Biotechnology Co., Ltd.) at different concentrations (25, 50 and 100 mg/ml) for 24 h at 37˚C. In addition, VSMCs were treated with ox-LDL (50 mg/ml) for different times (12, 24 and 48 h) at 37˚C.
5
Ox-LDL Induced VSMC Calcification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HA-VSMCs were purchased from the American Type Culture Collection (cat. no. CRL-1999) and cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37˚C with 5% CO2. Different concentrations of ox-LDL (Yeasen Biotechnology (Shanghai) Co., Ltd., Shanghai, China) dissolved into DMEM medium (0, 50, 100 and 150 µg/ml) were added to induce cell calcification at 37˚C for 48 or 72 h.
6
Silencing ZBTB20 in Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The siRNA targeting ZBTB20 was synthesized by the RiboBio company (Guangzhou, China), and the scrambled siRNA was served as the negative control (NC). For the cell transfections, the ZBTB20-siRNA or the scrambled siRNA was transfected into the macrophages with RNAiMAX (Thermo Fisher Scientific) [25 (link)]. Forty-eight hours later, the transfected macrophages were collected for further experimentation.
The ox-LDL was from Yeasen Biotech Co., Ltd., and a concentration of 50 μg/ml was adopted for respective time durations. After that, the macrophages were subjected to further experimental assays. The NRF2 inhibitor (ML385) was purchased from Selleck, and a concentration/duration of 5 μM/24 h was used before further experimental assays [26 (link)].
The ox-LDL was from Yeasen Biotech Co., Ltd., and a concentration of 50 μg/ml was adopted for respective time durations. After that, the macrophages were subjected to further experimental assays. The NRF2 inhibitor (ML385) was purchased from Selleck, and a concentration/duration of 5 μM/24 h was used before further experimental assays [26 (link)].
7
Ox-LDL-Induced Endothelial Injury Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HUVECs provided by Procell Co., Ltd., (Wuhan, China) were maintained in specific medium (Ham’s F-12K + 0.1 mg/mL heparin + 0.05 mg/mL endothelial cell growth supplement + 10% fetal bovine serum) (Procell Co., Ltd.,) at a 37℃ incubator containing 5% CO2.
Ox-LDL and ATV were purchased from Yeasen (Shanghai, China) and Solarbio (Beijing, China), respectively. To construct endothelial cell injury models for AS, HUVECs were treated with Ox-LDL at various doses (0, 25, 50, 100 and 200 mg/L). ATV was dissolved in ethanol (20 mg/mL), and Ox-LDL-treated HUVECs were administered with different concentrations of ATV (0, 2.5, 5 and 10 μM) for 24 h. Following experiments were performed using Ox-LDL at 100 mg/L and using ATV at 10 μM.
Ox-LDL and ATV were purchased from Yeasen (Shanghai, China) and Solarbio (Beijing, China), respectively. To construct endothelial cell injury models for AS, HUVECs were treated with Ox-LDL at various doses (0, 25, 50, 100 and 200 mg/L). ATV was dissolved in ethanol (20 mg/mL), and Ox-LDL-treated HUVECs were administered with different concentrations of ATV (0, 2.5, 5 and 10 μM) for 24 h. Following experiments were performed using Ox-LDL at 100 mg/L and using ATV at 10 μM.
8
Macrophage Differentiation and Foam Cell Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human monocytic cell line (THP-1) (ATCC) were grown in complete RPMI-1640 [20% FBS (AU0600); 1% Gluta-max; 1% sodium pyruvate]. Cells were cultured at 37°C in 5% CO2 and subcultured at 80%–90% confluence. We used a 100 ng/mL concentration of PMA (Merck) to induce macrophage formation. To induce foam cell formation, the macrophages were incubated with 25 or 50 µg/mL ox-LDL (Yeasen, 20605ES05, China) for 24 h. Foam cells were assessed by Oil Red O Staining kit (Beyotime, C0158S, China).
9
Transmigration Assay for Endothelial Dysfunction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The transmigration assay was performed as previously described [20] , with modifications. HUVECs were seeded onto the fibronectin coated (Sciencell) insert of a 24-well Transwell apparatus (6.5 mm, 8 μm pore size, Costar). HUVECs were cultured until confluent and then stimulated with 50 μg/mL ox-LDL (Yeasen) for 24 h. 1 × 10 5 U-937 cells were labelled with Calcein-AM (BioLegend) and added into the upper chamber. The number of cells that transmigrated to the bottom chamber in response to the conditioned medium or control medium were imaged using a confocal microscope (Leica TCS SP8X) and quantified by ImageJ software.
10
Generating Conditioned Medium from U-937 Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human monocytic U-937 cells were from the American Type Culture Collection (Manassas, VA, USA) and the THP-1 cells were purchased from the National Infrastructure of Cell Line Resource (Beijing, China). Cells were harvested in RPMI 1640 medium, supplemented with 10% foetal bovine serum and penicillin/streptomycin (Corning) at 37 °C with 5% CO 2 in a humidified atmosphere. Human umbilical vein endothelial cells (HUVECs) were obtained from ScienCell (San Diego, CA, USA) and maintained according to the vendor's instructions.
The conditioned medium was prepared as previously described [13] , with modifications. U-937 cells were differentiated in growth medium containing 100 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) for 48 h. U-937 macrophages were stimulated with 50 μg/mL Ox-LDL (Yeasen) for 24 h, and supernatant was collected as conditioned medium. Conditioned medium from unstimulated U-937 macrophages and empty RPMI-1640 medium were used as the control and basal media, respectively.
The conditioned medium was prepared as previously described [13] , with modifications. U-937 cells were differentiated in growth medium containing 100 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) for 48 h. U-937 macrophages were stimulated with 50 μg/mL Ox-LDL (Yeasen) for 24 h, and supernatant was collected as conditioned medium. Conditioned medium from unstimulated U-937 macrophages and empty RPMI-1640 medium were used as the control and basal media, respectively.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!