Lovastatin

Fluvastatin, pravastatin, lovastatin, and simvastatin were all obtained from Cayman Chemicals (Ann Arbor, MI); atorvastatin and rosuvastatin were obtained from Enzo Life Sciences, Inc. (Farmingdale, NY).

Pfizer Licensed Compound Library (81 drug compounds supplied as predissolved DMSO solutions at a concentration of 10 mM) was purchased from Selleck Chemicals. Food and Drug Administration (FDA)-approved drugs screening library (875 FDA-approved drug compounds supplied as predissolved DMSO solutions at a concentration of 10 mM), atorvastatin (calcium salt), bexarotene, adapalene, and lovastatin were from Cayman Chemical. Sypro Orange and ANS probes were from Sigma-Aldrich. Strain BL21-Gold (DE3) of Escherichia coli was obtained from Stratagene. Ni²⁺-nitrilotriacetic acid (Ni-NTA) resin was purchased from Thermo Fisher Scientific. 4-[¹⁴C]Cholesterol (0.1 mCi/ml, specific activity 50 mCi/mmol) was obtained from American Radiolabeled Chemicals. Other reagents and consumables were purchased from Sigma-Aldrich, Biochrom AG, Lonza, Thermo Fisher Scientific, Bio-Rad, Roche, or other standard commercial sources.

See Supplemental Text for details of cholesterol extraction. Ketoconazole and clotrimazole were obtained from the National Cancer Institute Developmental Therapeutics Program and Ro 48-8071 was from MedChem Express. Lovastatin and Simvastatin were obtained from Cayman Chemical.

Cell culture plasticware was purchased from Corning. All cell culture media and additives where purchased from Life Technologies except fetal bovine serum, which was purchased from Moregate Biotech. All-trans retinoic acid, uridine, 5-fluorodeoxyuridine and arabinofuranosyl were purchased from Sigma. The Aβ_1–42 and Aβ_42–1 fragments were purchased from Bachem and lovastatin and simvastain were purchased from Cayman Chemicals.

Lovastatin, fluvastatin sodium, simvastatin, atorvastatin, CoQ10, and mevalonic acid (MVA) were purchased from Cayman Chemical (Ann Arbor, MI, USA). 2-Deoxy-D-glucose (2-DG), carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), and oligomycin A were purchased from MedChem Express (Monmouth Junction, NJ, USA). Cholesterol-Water Soluble, FPP, geranylgeranyl pyrophosphate (GGPP), trichloroacetic acid (TCA), CHX, MG132, DNase-free RNase A, triton X-100, crystal violet, and sulforhodamine B (SRB) were obtained from Sigma Aldrich (St. Louis, MO, USA). Antibody sources were as follows: HMGCR (BD Biosciences, San Jose, CA, USA); PKM2 (Cell Signaling, Danvers, MA, USA); Hsp90 (Abcam, Cambridge, Cambridgeshire, UK), β-actin (Sigma-Aldrich, St. Louis, MO, USA); horseradish peroxidase-conjugated secondary antibodies (Jackson Laboratory, Bar Harbor, ME, USA).

MSCs were starved for 15-18 hours in serum-free DMEM/LG containing 0.5% fetal bovine serum (Thermo, Logan, UT), then treated with 2 μg/ml IL-1β inhibitor IL-1RA (PeproTech, NJ, USA) for 2 hours prior to cytokine stimulation. The LFA-1/ICAM-1 inhibitor lovastatin (Cayman Chemical, USA) was added to the cell coculture at a concentration of 50 μM. MAPK p38 inhibitor SB 203580 (5 μM) (Tocris, UK) was added to the cell culture after IL-1β stimulation. According to our previous study, MSCs treated with 100 ng/ml human recombinant IL-1β for 18 hours significantly enhanced migration without affecting cell viability and cell proliferation [10 (link), 32 (link)]. At the indicated time, cells were incubated for 30 minutes with 100 ng/ml human recombinant IL-1β (PeproTech, NJ, USA) in the continued presence of these inhibitors.
HUVECs were starved for 3 hours in serum-free DMEM/F12 containing 1% bovine serum albumin (Sigma, MO, USA), then treated with 100 ng/ml IL-1β for 6 hours.