Fig. 2

Knockdown or pharmacological inhibition of G6PD protected DA neurons from LPS-elicited inflammatory insults. a Primary mouse microglia-enriched cultures were transfected with 10 nM scramble RNA (SS) or G6PD siRNAs (GS1 and GS2) for 30 h before examining the knockdown efficiency by Western blotting assay. b–d The reconstituted cultures, which were prepared by adding mouse microglia (5 × 10⁴ microglia/well) with 30-h knockdown of G6PD by siRNAs onto mouse neuron-astrocyte layer, were treated with vehicle or LPS for 5–6 days. Western blotting assay (b), representative images (c), and cell counting (d) from three to four experiments revealed DA neuroprotection by knockdown of microglial G6PD. e–j Rat mesencephalic neuron-glia cultures were pretreated with vehicle, 6-AN, or DHEA for 30 min and treated with the vehicle or LPS. e Effects of 6-AN and DHEA on G6PD enzyme activity and NADPH levels at 2 days after LPS treatment. f–j Survival of DA neurons was determined by ³[H]DA uptake assay (f, g), western blotting assay (h, i), and representative images (j) at 7 days after LPS treatment. Results are mean ± SEM (a, d–g, i). *p < 0.05 compared with vehicle-treated controls. ^#p < 0.05 compared with LPS-treated cultures