Fig. 6.

Microglia proinflammatory mediator release in response to NT is attenuated by the PI3K/mTOR inhibitors and the flavonoids Lut and Methlut. HM-SV40 (5 × 10⁴ cells) were pretreated with the dual PI3K/mTOR (PF, 0.1 μM) and the mTOR (Rap and KU, 0.1 μM) inhibitors or flavonoids [Lut and Methlut, 5 μM (Upper) or 0.1–10 μM (Lower)] for 2 h, and then stimulated with NT (10 nM) for 24 h in serum-free medium to measure release of IL-1β (A), CXCL8 (B), CCL2 (C), and CCL5 (D) by ELISA. All inhibitors were dissolved in water or DMSO with a final concentration <0.1%. All conditions were performed in triplicate for each dataset and were repeated three times (n = 3). Significance of comparisons was determined for stimulated cells and for those cells with inhibitors/flavonoids, as denoted by the horizontal lines (P < 0.001 or P < 0.0001), and also among each of the inhibitor/flavonoid treatments shown by the horizontal brackets and by corresponding *P < 0.05, **P < 0.001, and ***P < 0.0001.