The largest database of trusted experimental protocols

4 protocols using MBMECs

Mouse brain microvascular endothelial cells (mBMECs, from Angio-Proteomie) were cultured in DMEM/HG medium (ThermoFisher, Waltham, MA, USA) supplemented with 20% FBS (ThermoFisher, Waltham, MA, USA), and 1% Antibiotic–Antimycotic (100×) solution (ThermoFisher, Waltham, MA, USA). Cells were infected with lentivirus carrying three short hairpin RNA (shRNA) sequences against mouse Sirt3 or a scramble lentivirus and selected with puromycin for stable expression. Puromycin was added 3 days after infection, with a concentration of 1 μg/mL, which was replaced every 2–4 days depending on the confluency of the cells. Human umbilical vein endothelial cells (HUVECs, from Lonza, Visp, Switzerland) between passages 6–9 were cultured in DMEM/F12 medium supplemented with 10% FBS, 1% Antibiotics–Antimycotics, and endothelial cell growth supplement (Sigma-Aldrich, St. Louis, MO, USA). HUVECs were transfected with human SIRT3 siRNA (GenePharma, Suzhou, China) or universal scrambled negative control siRNA using Lipofectamine RNAiMAX Transfection Reagent (ThermoFisher, Waltham, MA, USA), according to the manufacturer’s instructions and were used within 36–48 h after transfection.
+ Open protocol
+ Expand
Mouse brain microvascular endothelial cells (mBMECs) (Angio-Proteomie, Boston, MA, USA) were cultured in DMEM (Gibco) with 4.5 g/L glucose, 0.5% FBS (Gibco), 100 IU/mL penicillin and 100 μg/mL streptomycin (Penicillin-Streptomycin, Gibco) at 37 °C in a CO2 incubator, divided into groups of control, 1 pg/mL IL-1β and 1 pg/mL IL-1β + 0.2 μg/mL CBSF.
+ Open protocol
+ Expand
Mouse brain microvascular endothelial cells (mBMECs) (Angio-Proteomie, Boston, MA, USA) were cultured in DMEM (Gibco) with 4.5 g/L glucose, 10% fetal bovine serum (FBS Gibco) and 100 IU/mL penicillin and 100 μg/mL streptomycin (Penicillin-Streptomycin, Gibco), with or without IL-1β (0.5 ng/mL) treatment, PCA (1–100 nM) and Akt inhibitor IV (1 μM) overnight at 37°C in a CO2 incubator.
+ Open protocol
+ Expand
Mouse brain microvascular endothelial cells (mBMECs) (Angio-Proteomie, Boston, MA, USA) were cultured in DMEM (Gibco) with 4.5 g/L glucose, 0.5% FBS (Gibco), 100 IU/mL penicillin and 100 μg/mL streptomycin (Penicillin-Streptomycin, Gibco) at 37 °C in a CO 2 incubator, divided into groups of control, 1 pg/mL IL-1β and 1 pg/ mL IL-1β + 0.2 μg/mL CBSF.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!