Additionally, cytokine profile (IL-1β, IFN-γ, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, and TNF-α) using multiplex cytokine assay (Thermo Fisher Scientific, Waltham, MA, USA) as described [30] was performed in collected media supernatant from insert (luminal) and 24-well plate (abluminal) compartments prior to hBMEC fixation after 5 DIV of exposure to 10% FBS, plasma from ALS or control patient. Here, TNF is linked to amyotrophic lateral sclerosis.