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. 2014 Oct 20;21(12):1741–1758. doi: 10.1089/ars.2013.5587

FIG. 5.

FIG. 5.

Knockdown of AMPK by siRNA attenuated ADT-OH promoting effects on M2 polarization of BV2 microglial cells. (A) AMPK siRNA significantly reduced AMPK protein expression in BV2 cells (n=3). * p<0.05, compared with nontransfected control cells (Con) or cells transfected with nonsense siRNA (NC). (B–E) qPCR measurement of mRNA expression of M1 genes iNOS (n=4), TNF-α (n=3), IL-1β (n=4), and IL-6 (n=3) in BV2 cells at 6 h after LPS stimulation. ADT-OH suppression of LPS-evoked M1 gene expression (iNOS, TNF-α, IL-1β, and IL-6) was markedly attenuated by the AMPK siRNA. (F, G) qPCR measurement of mRNA expression of M2 genes arginase 1 (n=5) and YM1/2 (n=4) in BV2 cells at 6 h after LPS stimulation. ADT-OH-enhanced mRNA expression of arginase 1 and YM1/2 was markedly attenuated by the AMPK siRNA. (H) ELISA measurement of IL-10 protein levels in media of BV2 cell cultures at 24 h after LPS stimulation (n=4). ADT-OH elevated IL-10 production was markedly attenuated by the AMPK siRNA. *p<0.05 or **p<0.01, compared with control cells treated with vehicle (Con); #p<0.05 or ##p<0.01, compared with cells treated with LPS alone (LPS); and $p<0.05 or $$p<0.01, compared with nontransfected cells treated with LPS plus ADT-OH (LPS+ADT-OH) or NC-transfected cells treated with LPS plus ADT-OH (LPS+ADT-OH+NC).