Hbmec

hBMECs were purchased from Neuromics (Catalog code number: HEC02; Minneapolis, MN, USA). HUVECs were purchased from ThermoFisher Scientific (Catalog code number: C0035C; Waltham, MA, USA). Cells were cultured at early passages (3–7) under standard conditions (37 °C, 5% CO₂), as previously described [48 (link)]. In some assays, the cells were transfected with pcDNA3.1-FGFBP1 plasmids obtained from GenScript (Piscataway, NJ, USA). All other reagents were obtained from Merck (Darmstadt, Germany).

All reagents were purchased from Millipore-Sigma (Burlington, MA, USA), unless otherwise stated. We obtained hBMECs from Neuromics (Minneapolis, MN, USA; #HEC02) [111 (link)]; these cells have been proved to be the most suitable human cell line to reproduce the BBB in vitro [112 (link)]. Adult human lung microvascular endothelial cells (HMVEC-Ls) were obtained from Lonza (Basel, Switzerland; catalog number, CC-2527) and human umbilical vein endothelial cells (HUVECs) from ThermoFisher Scientific (Waltham, MA, USA; Catalog number, #C0035C). Cells were cultured in a standard humidified atmosphere (37 °C) containing 5% CO₂, as we previously described [111 (link),113 (link)]. In some experiments, cells were transfected with pcDNA3.1-TIM-1 plasmids (GenScript, Piscataway, NJ, USA).

All reagents were purchased from Millipore-Sigma (Burlington, MA, USA), unless otherwise stated. Human brain microvascular endothelial cells (hBMECs) were obtained from Neuromics (Minneapolis, MN; catalog number: #HEC02). These cells have been proved to be the most suitable human cell line for an in vitro blood–brain barrier (BBB) model [29 (link)].
Cells were cultured in a standard humidified atmosphere (37°C) containing 5% CO₂. In some experiments, cells were transfected with pcDNA3.1-Neuropilin-1 plasmids (GenScript, Piscataway, NJ).

All reagents were purchased from Millipore-Sigma (Burlington, MA, USA), unless otherwise stated. Human brain microvascular endothelial cells (hBMECs) were obtained from Neuromics (Minneapolis, MN, USA; catalog number: #HEC02). These cells have been proved to be the most suitable human cell line for an in vitro blood–brain barrier (BBB) model [29 (link)].
Cells were cultured in a standard humidified atmosphere (37 °C) containing 5% CO₂. In some experiments, cells were transfected with pcDNA3.1-Neuropilin-1 plasmids (GenScript, Piscataway, NJ, USA).

Mouse Brain Endothelial Cells (BEND3) and Human Brain Microvascular Endothelial Cells (HBMECs) were obtained from American Type Culture Collection (Cat # CRL-2299, Manassas, VA, United States) and NEUROMICS (Cat # HEC02, Edina, MN, United States), respectively. BV2 cells, mouse microglial cell line, cells were a kind gift from Dr. Dennis Selkoe (Harvard Medical School, Boston, MA, United States). BEND3 and BV2 cells were cultured in advanced DMEM/F12 medium (Cat # 12634-010, Gibco, Gaithersburg, MD, United States) containing 10% fetal bovine serum and 1% penicillin-streptomycin under standard culture conditions, and Endo-Growth Media (Cat # MED001, NEUROMICS, Edina, MN, United States) was used to grow HBMECs. Generation of Sirt3-silenced cell lines: BEND3 and BV2 cells were infected with Sirt3 shRNA Lentiviral particles (Cat # sc 61556) or control shRNA Lentiviral particles (Cat # sc 108080) using polybrene (0.5 μg/ml; Cat # sc 134220). Stable infected cells were selected using puromycin dihydrochloride (8 μg/ml, Cat # sc 10807) in DMEM media supplemented with 10% FBS for 4 weeks, and cells were pooled and maintained in the same medium. All the reagents, used for transfection, were from Santa Cruz Biotechnology (Dallas, TX, United States). Silencing of Sirt3 gene was confirmed by western blot analysis in stable infected cells.

Unless otherwise stated, the reagents used in this study were obtained from Millipore Sigma, Burlington, Massachusetts, USA. Human brain microvascular endothelial cells (HBMECs) were obtained from Neuromics (Minneapolis, Minnesota, USA) and cultured at 37°C with 5% CO₂.