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. 2014 Feb 11;9(2):e87030. doi: 10.1371/journal.pone.0087030

Figure 7. Effects of 4d and 4e on mRNA expressions of proinflammatory molecules in LPS-stimulated BV2 microglial cells.

Figure 7

(A) Effect of 4d and 4e on mRNA expression of iNOS, COX-2 and pro-inflammatory cytokines in LPS-stimulated BV2 cells. Cell were pre-treated with 4d or 4e (5 µM) for 1 h prior to LPS stimulation. Total RNA was isolated after 6 h of LPS treatment. The mRNA levels for iNOS, TNF-α, IL-1β, IL-6 and COX-2 were measured by quantitative real-time PCR. The gene expression was normalized by GAPDH expression. Data are represented as mean ± S.E.M. of three independent experiments. *P<0.05; significantly different from LPS-treated cells. (B) BV2 cells were pre-treated with 4d or 4e (5 µM) for 1 h, and then stimulated with LPS for 3 h. Subsequently, 10 µg/ml actinomycin D (Act D) was added. After 0 min, 15 min, 30 min, and 1 h, cells were harvested and TNF-α mRNA expression was quantified by real-time PCR. The graphs shows mean TNF-α mRNA normalized to GAPDH mRNA expressed as a percentage of t = 0 Act D treatment ± S.E.M. of three independent experiments.