Primary HMVEC from male and female cadavers were purchased (Lonza, Basel, Switzerland) and used at passage 3–6. HMVEC were plated and grown to 90–100% confluence before treatment (at 37°C, 5% Co2), using the appropriate medium (endothelial cell growth medium [EGM]–2) (Lonza). Cells were incubated with lipopolysaccharide (LPS) (1 μg/mL; Sigma-Aldrich, St. Louis, MO), tripalmitoyl-S-glyceryl cysteine (Pam3Cys, 10 μg/mL; Abcam, Cambridge, United Kingdom), polyinosinic:polycytidylic acid (10 μg/mL; Abcam), recombinant human interleukin (IL)–1β (10 ng/mL; R&D Systems, Minneapolis, MN), recombinant human tumor necrosis factor (TNF)–α (10 ng/mL; R&D Systems), or cytomix (TNF-α, interferon-γ, and IL-1-β; R&D Systems). Simultaneously or after 8 hours, cells were stimulated with epinephrine (Sigma-Aldrich), norepinephrine (Hospira, Lake Forest, IL), vasopressin (Anaspect, Fremont, CA), or angiotensin-2 (Sigma-Aldrich) from 0.1 to 100 μM, encompassing clinically relevant concentration. The following reagents have been used to decipher the involvement of different adrenergic receptors (ARs): dobutamine, atenolol, propranolol (Cerilliant, Round Rock, TX), ICI-118,551 (Tocris, Bristol, United Kingdom), phentolamine methanesulfonate salt (PMS; Sigma-Aldrich), and phenylephrine (Hikma, Eatontown, NJ). UCSF’s Institutional Review Board waived the need for approval to work with deidentified human cells purchased from a vendor.

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