HK-2 cells (TCH-C400) were obtained from Haixing Biosciences (Suzhou, China) and cultured in high glucose DMEM containing 10% FBS and 1% penicillin‒streptomycin. The cells were maintained at 37 °C in a 5% CO2 atmosphere. Briefly, HK2 cells were firstly cultured for 24 h to reach about 80% confluence. New DMEM without FBS were then used for HK-2 starvation overnight before being treated with TGF-β1 (10 ng/ml, ABclonal, China) to induce the EMT process, which mimics the process in kidney fibrosis. The cells in some experiments were pretreated with the Carnitine palmitoyltransferase 1 A (CPT1A) activator C75 (10 µM, HY-12,364, MedChemExpress, USA) or the CPT1A antagonist Etomoxir sodium salt (80 µM, HY-50,202 A, MedChemExpress, USA) for 2 h [28 (link)].
Hy 12364
Manufactured by MedChemExpress
Sourced in United States
HY-12364 is a laboratory equipment product offered by MedChemExpress. It is designed for general laboratory use.
Automatically generated - may contain errors
Lab products found in correlation
Tgf β1, by ABclonal (1 mentions)
L2880, by Merck Group (1 mentions)
D12492, by Research Diets (1 mentions)
Orlistat, by Merck Group (1 mentions)
Hy n1966, by MedChemExpress (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Hy 50202, by MedChemExpress (1 mentions)
Etomoxir, by MedChemExpress (1 mentions)
5 protocols using hy 12364
1
CPT1A Modulates TGF-β1-Induced EMT in HK-2 Cells
2
FASN Regulation in Clear Cell Renal Carcinoma
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Increasing the expression level of fatty acid synthase (FASN) was positively correlated with aggressive cell proliferation, migration, apoptosis, and lipid droplet formation. It was also shown to regulate metabolic disorders associated with clear cell renal carcinoma. Researchers also proved that pharmacological inhibitor of FANS could suppress the growth and invasiveness of clear cell renal carcinoma. Therefore, we used two human clear cell renal carcinoma cell line (caki-1 and 786-0 cell lines) from authenticated cell cultures of the Chinese national collection. We down-regulated the expression of FANS in caik-1 cells by the way of siRNA virus and exposed FASN inhibitorC75 (HY-12364, MedChemExpress, China) with two cell line (caki-1 and 786-0). We verified the expression level of survival-related fatty acid genes with quantitative real-time polymerase chain reaction (Q-PCR) [16 (link)].
3
Dietary-induced Obesity Mouse Model of ALI
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Six-week-old mice were randomly divided into a regular chow diet (Beijing Keao Xieli Feed Co., Ltd, 12% kcal fat) group (lean) or a high-fat diet (Research Diets, D12492, 60% kcal fat) group [diet-induced obese (DIO)]. After feeding for 24 weeks, the mouse model of ALI was established according to the previous literature [21 (link)]. Briefly, mice were anesthetized with pentobarbital and LPS (O55:B5, L2880, Sigma, 100 μg dissolved in 50 μL of saline) or vehicle (50 μL of saline) was instilled intratracheally into both lean and DIO mice. For FASN inhibition, DIO mice were intraperitoneally injected with C75 [MedChemExpress, HY-12364, dissolved in 10% dimethyl sulfoxide (DMSO)] at 10 mg/kg body weight at 30 min before LPS administration, and mice in other group were intraperitoneally injected with DMSO (vehicle) at the same concentration. Mice were euthanized and sacrificed at 6 h or 8 h after LPS administration, and their lung tissues were harvested for subsequent procedures.
4
Inhibition of FASN in Colorectal Cancer
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For the application of the FASN synthetase inhibitor Orlistat (O4139-25MG, Merck, Darmstadt, Germany) in vitro, the inhibitor was prepared by referencing the procedure of Browne et al. [17 (link)], and 200 µmol/L Orlistat was adopted to treat CRC cells for 48 h when necessary. Another FASN inhibitor, C75 (25–200 µmol/L) (HY-12364, MedChemExpress, Monmouth Junction, NJ, USA), was used to treat CRC cells for 48 h as per a report from Chen et al. [11 (link)]. Based on a previous study [18 (link)], 800 µmol/L of palmitoleic acid (HY-N1966, MedChemExpress) was applied for 72 h for in vitro experiments.
5
In Vitro Decidualization of Human Endometrial Stromal Cells
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Human ESCs were a gift from Prof. Wang Haibin of Xiamen University. Endometrial stromal cells were grown in DMEM/F12 medium containing 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin, and cultured in an incubator at 37 °C with 5% CO2. For the in vitro induced ESC decidualization model, ESCs were cultured in DMEM/F12 containing 2% FBS and treated with medroxyprogesterone acetate (MPA; 1 μM; #B1510; APExBIO Technology, Houston, TX, USA) and dibutyryl-cAMP (db-cAMP; 0.5 mM; #B9001, APExBIO) for 3, 5, and 7 days. The induction medium was changed every 2 days. Endometrial stromal cells induced to decidualization in vitro were treated with the carnitine palmitoyl transferase (CPT)1 inhibitor, etomoxir (25 μM, 50 μM; #HY-50202; MedChemExpress, Monmouth Junction, NJ, USA) for 24 h or 48 h; the diacylglycerol acyltransferase 2 (DGAT2) inhibitor, PF-06424439 (10 μM, 20 μM; #HY-108341A; MedChemExpress) for 24 h or 48 h; the DGAT1 inhibitor, PF-04620110 (5uM; #HY-13009; MedChemExpress) for 24 h; and CPT1 agonists, baicalin (25 μM, 50 μM; #HY-N0197; MedChemExpress), and C75 (1 μM, 4 μM; HY-12364, MedChemExpress) for 24 h, respectively. For siRNA experiments, 100 nM siRNA was transfected into cells using 5 μL Lipofectamine 2000 (#11668019, Invitrogen) and the cells were collected after 48 h. The interference sequences for targeted genes are shown in Supplementary Table 3 .
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