Fig. 5

Inhibition or knockdown of G6PD suppressed NF-кB activation. a Microglia-enriched cultures were challenged by LPS for 15, 30, and 60 min. The activation of NF-кB pathway, as shown by phosphorylation and nuclear translocation of p65, was observed after LPS treatment. b–d Microglia-enriched cultures were pretreated with vehicle, 6-AN (10 μM), or DHEA (100 μM) for 30 min prior to the addition of LPS. b Immunofluorescence was performed with phosphorylated p65 (red) 30 min after LPS treatment. c, d Nuclear translocation of p-p65 and p65 was examined after the cultures were treated with 6-AN or DHEA. e–g At 30 h after transfection with scramble siRNA (SS) and G6PD siRNAs (GS1 and GS2), mouse microglia-enriched cultures were challenged with vehicle or LPS. Nuclear and cytoplasmic proteins were examined at 30 min after LPS treatment to determine the effect of reduced G6PD on NF-кB activation. Results are mean ± SEM of two to three experiments performed in triplicate (a, d, f). *p < 0.05 compared with vehicle-treated controls. ^#p < 0.05 compared with LPS-treated cultures