Nabut

All experiments were performed using a robotic pipetting workstation (Hamilton Robotics Star Line) for high seeding, stimulation and revelation homogeneity. For each experiment, cells were seeded at 25000 cells/well in 96 well-plates and incubated 24 h prior to activation. Homogeneous cellular activations were performed using sodium butyrate (NaBut, 2 mM, Sigma) for PPARγ and ANGPTL4 reporter systems or 0,45 µm-filtered, sonicated E. coli (Epi300 from Epicentre, 10% vol/vol) for NFκB-SEAP system. Cells were then incubated 24 h prior quantification of the reporter gene expression (alkaline phosphatase (SEAP) or luciferase). SEAP (for NFκB system) secretion or intracellular luciferase expression (for both ANGPTL4 and PPARγ systems) were revealed using the QUANTI-Blue (Invivogen) or One-Glo (Promega) reagent respectively, according to the manufacturers’ instructions. All measurements were performed using a microplate reader (Infinite 200, Tecan) and quantified at 655 nm OD for SEAP and by luminescence for luciferase.

α-Amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA; Abcam) was infused through an Alzet osmotic minipump (Model 2004; capacity, ∼234 µl; flow rate, 0.23 μl/h) directly into the spinal cord tissue. Osmotic minipumps were filled with PBS (for control groups) or with 1.5 mM (low-dose) and 3.0 mM (high-dose) AMPA dissolved in filtered PBS, and were incubated in filtered isotonic saline solution at 37°C for 48 h before surgical implantation to achieve a constant flow rate. NaBut (Sigma-Aldrich) at a dose of 500 mg/kg (0.9 M), or PBS (as control), was administered intraperitoneally. Each batch of NaBut solution was freshly prepared in sterile filtered PBS. Intraperitoneal administration was performed daily after motor behavior assessment for 6 days or 9 days after minipump surgical implantation (see below). The combination of osmotic minipump infusion and i.p. administration yielded six groups: control (vehicle spinally infused and vehicle administered intraperitoneally), control+NaBut, AMPA low-dose or AMPA high-dose+vehicle, and AMPA low-dose or AMPA high-dose+NaBut. Every AMPA low-dose or high-dose group was surgically implanted and studied at the same time as the other groups.

5‐AzadC, NaBut, VPA, and SAHA (vorinostat) were purchased from Sigma Aldrich. MS‐275 was purchased from Enzo Life Sciences. Belinostat, panobinostat, and romidepsin were purchased from Selleckchem.