VSMCs were obtained from Keygen Biotech. The cells were cultured in DMEM containing 10% foetal bovine serum and maintained at 37°C in 5% CO2 environment with medium changed every 2 days. For HMGB‐1 induction experiment, the cells were exposed to AGEs (100 μg/mL) for 0, 6, 12 and 24 hours. Then, 100 μg/mL AGEs for 24 hours were used in the following experiments. For the in vitro calcification experiments, VSMCs were incubated with β‐glycerophosphate (β‐GP, 10 mmol/L) in medium containing HMGB‐1 (100, 200, 400 μg/mL) or TM (1, 2, 4 mg/L) for two weeks. Then, 200 μg/mL HMGB‐1 was used in the following experiments. For the inhibitory studies, 10 mmol 4‐PBA or 100 μmol/L Gly was pre‐treated for 2 hours before coincubation with AGEs or HMGB‐1.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.