Fig. 8.

Inhibition of neuroinflammation by thymoquinone was abolished in the presence of EX527. BV2 cells were treated with EX527 (1 µM), followed by thymoquinone (10 µM) and LPS (100 ng/ml) for 24 h. Culture supernatants were collected and analysed for TNFα (a), IL-6 (b), IL-1β (c), nitrite (d) and PGE₂ (e). In f nuclear extracts from cells were added to 96-well plates to which an oligonucleotide containing the NF-κB consensus site (5′-GGGACTTTCC-3′) has been immobilised, followed by addition of NF-κB and HRP-conjugated antibodies. Absorbance was read in a microplate reader. Reduction of cellular ROS production by thymoquinone was not abolished in the presence of EX527 (G). (Mean ± SEM; **p < 0.01, ***p < 0.001 compared with LPS stimulation; ^# p < 0.05, ^## p < 0.01, ^### p < 0.001, EX527 + thymoquinone + LPS treatment compared with thymoquinone + LPS treatment; one-way ANOVA)