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. Author manuscript; available in PMC: 2014 Jul 25.
Published in final edited form as: J Biomol Screen. 2013 Aug 14;19(1):17–31. doi: 10.1177/1087057113499406

Figure 3.

Figure 3

Modulation of tumor necrosis factor–α (TNF-α) release from microglial cell lines. (A) BV-2 cells and (B) C8-B4 cells (3 × 104 cells/well plated 16 h before experiment in poly D-Lys–coated 96-well plates) were treated with lipopolysaccharide (LPS) (1–1000 ng/mL) for 4 h in 2% fetal bovine serum (FBS) containing RPMI and Dulbecco's modified Eagle's medium, respectively. Media were removed from plates (25 μL) and used to determine levels of TNF-α released by the cells in the absence or presence of 20 μM microglia activation inhibitors apigenin and fisetin added to cells 30 min prior to LPS treatment. (C) Inhibition of microglial activation by the glutaminase inhibitor DON added to the cells 30 min prior to LPS treatment (10 ng/mL for 4 h). Levels of TNF-α were determined using an eBioscience (San Diego, CA) mouse TNF-α enzyme-linked immunosorbent assay kit (catalog number 88-7324-88) according to the manufacturer's instructions.