Fig. 9
Suppressing LPA1 activity attenuates LPS-upregulated mRNA expression levels of proinflammatory cytokines in mouse primary microglia. Primary microglia were stimulated with LPS (100 ng/ml). Thirty minutes prior to LPS exposure, cells were treated with AM095 (2 μM in 0.1% DMSO). Alternatively, primary microglia were stimulated with LPS at 2 days after transfection with lentivirus particles bearing LPA1 shRNA (shLPA1) or non-target control shRNA (shNC). Total RNA was extracted at 24 h after LPS exposure, and mRNA expression levels of proinflammatory cytokines were determined by qRT-PCR analysis. a–c Changes in mRNA expression levels of TNF-α, IL-6, and IL-1β. n = 3 per group. ***p < 0.001 versus the vehicle-treated cells. ##p < 0.01 and ###p < 0.001 versus LPS-stimulated cells. d–g Effects of LPA1 knockdown (d) on the mRNA expression levels of TNF-α, IL-6, and IL-1β (e–g). n = 3 per group. **p < 0.01 versus cells infected with non-target control shRNA lentivirus (shNC) in d. ***p < 0.001 versus vehicle-treated cells in e–g. ##p < 0.01 and ###p < 0.001 versus LPS-stimulated cells infected with non-target control shRNA lentivirus (shNC+LPS) in e–g