BMSCs were induced and cultured in osteogenic medium according to a StemPro Osteogenesis Differentiation Kit (Invitrogen, USA). Recombinant human BMP-4 protein (rhBMP-4) (R&D Systems, USA), a Smad inhibitor (LDN-193189) (Selleck, USA), and metformin (TargetMol, USA) were separately added to 0.5 ml medium in 24-well dishes (Corning, USA), and the final concentrations of each substance in the medium were as follows: rhBMP-4 (+: 10 ng/ml; ++: 50 ng/ml), LDN-193189 (+: 0.1 μM; ++: 0.5 μM), and metformin (+: 30 μM; ++: 100 μM). After 10 d of induction, cells were fixed in 70% ethanol for 1 h and stained using an ALP staining kit (Beyotime, China) according to the manufacturer's protocol. Intracellular ALP activity assays of BMSCs were performed at 3, 5, and 7 d of induction using an ALP Activity Assay Kit (Nanjing Jiancheng Bioengineering Institute, China) according to the manufacturer's protocol and were standardized based on protein concentration. After induction for 21 d, cells were fixed in 70% ethanol and stained with 2% alizarin red S staining solution (Sigma-Aldrich, USA) for 5 min. Then, 1 ml of isopropanol was added into each well to dissolve the red perylenequinone derivatives in the calcium nodules, and the optical density (OD) values were measured at a wavelength of 550 nm.
Metformin
Manufactured by Targetmol
Sourced in United States, China
Metformin is a chemical compound used in laboratory settings. It is a white crystalline powder that is soluble in water. Metformin is commonly used as a reference standard or analytical reagent in various scientific experiments and analyses.
Automatically generated - may contain errors
Lab products found in correlation
24 well dishes, by Corning (1 mentions)
Stempro osteogenesis differentiation kit, by Thermo Fisher Scientific (1 mentions)
Rhbmp 4, by R&D Systems (1 mentions)
Alizarin red s staining solution, by Merck Group (1 mentions)
Alp staining kit, by Beyotime (1 mentions)
Ldn193189, by Selleck Chemicals (1 mentions)
Alp activity assay kit, by Nanjing Jiancheng (1 mentions)
Simvastatin, by Targetmol (1 mentions)
Rosuvastatin, by Targetmol (1 mentions)
Empagliflozin, by Targetmol (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Sangon (1 mentions)
Ox ldl, by Solarbio (1 mentions)
Doxycycline, by Cayman Chemical (1 mentions)
Nifedipine, by Cayman Chemical (1 mentions)
Metformin, by Cayman Chemical (1 mentions)
Ha tag, by Merck Group (1 mentions)
Doxorubicin hydrochloride d1515, by Merck Group (1 mentions)
Flag tag 20543 1 ap, by Proteintech (1 mentions)
4 protocols using metformin
1
Osteogenic Differentiation of BMSCs
2
Evaluating Cardiovascular Pharmacotherapies
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TNF-α (#300-01A, PEPROTECH, Rocky Hill, USA); IL-1β (#200-01B; PEPROTECH, Rocky Hill, USA); oxLDL (#H7980, Solarbio, Beijing, China); DMSO (#A100231, Sangon Biotech, Shanghai, China); Atorvastatin (#222412-82-0, Cayman Biochem, Ann Arbor, Michigan, USA); Simvastatin (#79902-63-9, TargetMol, Shanghai, China); Rosuvastatin (#287714-41-4, TargetMol, Shanghai, China); Fenofibrate (#49562-28-9,TargetMol, Shanghai, China); Empagliflozin (#864070-44-0,TargetMol, Shanghai, China); Troglitazone (#97322-87-7, MCE, Shanghai, China); Metformin (#1115-70-4,TargetMol, Shanghai, China); Cilostazol (#73963-72-1,TargetMol, Shanghai, China); Fluvoxamine (#61718-82-9,TargetMol, Shanghai, China); Riociguat (#625115-55-1,TargetMol, Shanghai, China); Sildenafil citrate (#171599-83-0,TargetMol, Shanghai, China).
3
Zebrafish Pharmacological Compound Screening
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Concentrated stock solutions for pentylenetetrazole (PTZ, Sigma), metformin (Cayman Chemical), doxycycline (Cayman Chemical), nifedipine (Cayman Chemical), and pyrantel tartrate (TargetMol) were made by dissolving each in either ddH2O (PTZ, doxycycline, metformin) or DMSO (nifedipine, pyrantel tartrate). 7 dpf (or 5 dpf, in the case of the doxycycline treatment experiments) zebrafish larvae were placed individually into separate wells of a 96‐well plate in fish water. All water in each well was carefully drawn off, and 250 μL 1X Ringer's Solution (116 mmol/L NaCl, 2.9 mmol/L KCl, 1.8 mmol/L CaCl2, and 5 mmol/L HEPES) was pipetted into the well. Lastly, small volumes of concentrated stock solutions of drug (or vehicle) were delivered to their respective groups to reach their final concentrations: 5 mmol/L PTZ, 2 μmol/L metformin, 10 μmol/L doxycycline, 12 μmol/L nifedipine, and 75 μmol/L pyrantel tartrate. Solutions were mixed by gently tapping the plate. The larvae were divided such that all five groups had approximately equal representation on every plate (from 10 to 20 embryos per treatment group per plate).
4
Antibody and Drug Inventory for Cell Signaling Research
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The following antibodies were purchased from Cell Signaling Technology (America): SUFU (C81H7), LATS1 (3477S), P-LATS1 (9157S), P-YAP127 (4911S), V5-tag (13202S), Myc-tag (2278S), xCT/SLC7A11 (12691). Anti-V5 (P/N46-0705) was obtained from Invitrogen Thermo Fisher Scientific (America). YAP1 (NB110-58358) was purchased from NOVUS (America). GAPDH (10494-1-AP), AXL (13196-1-AP) and Flag-tag (20543-1-AP) were bought from Proteintech (America). HA-tag (H6908) was purchased from Sigma (America). LAMIN A + C (ab133256), ACSL4 (ab155282), GPX4 (ab125066) and Ki67 (ab16667) were bought from Abcam (UK). SUFU (sc-137014) and IgG (sc-69786) were purchased from Santa Cruz Biotechnology (America). Verteporfin (S1786), Z-VAD-FMK (S7023) and Necrostatin-1 (S8037) were purchased from Selleck Chemicals (America). Ferrostatin-1 (SML0583), liproxstatin-1 (SML1414), 1S,3R-RSL3 (SML2234) and Doxorubicin Hydrochloride (D1515) were obtained from Sigma (America). 3-Methyladenine (189490) was bought from CALBIOCHEM (Germany). Metformin (T0740) was purchased from TargetMol (America). Vincristine (594515) was bought from J&K Scientific (China). 5-Fluorouracil (51-21-8) was obtained from Aladdin (China). Paclitaxel (SP8020) and Cisplatin (SC5170) were purchased from Solarbio (China). Drugs were dissolved and stored at − 20°C or −80°C according to the manufacturer’s instructions.
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