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Figure 3 | Molecular Pain

Figure 3

From: Mitogen activated protein kinase phosphatase-1 prevents the development of tactile sensitivity in a rodent model of neuropathic pain

Figure 3

Reversal effects of BV-2 cell MKP-1 induction in cytokine production by siRNA MKP-1 knockdown. (a) Efficient siRNA BV-2 cell transfection using Alexa-555 red fluorescent-labeled siRNA. (b) MKP-1 mRNA level in BV-2 cells stably expressing MKP-1 after 24 h transfection with MKP-1 siRNA compared to negative control siRNA and vehicle treated cells. IL-6 (c), TNF-α (d), and IL-1β (e) mRNA quantification (qRT-PCR) in control (normal) and stably expressing MKP-1 BV-2 cells transfected 24 h with a negative siRNA, or a siRNA against MKP-1 and challenged with LPS (1 μg/ml) for 6 h (IL-6 and TNF-α) or 12 h (IL-1β). Since MKP-1 siRNA at 10 nM displayed an opposite effect than expected in TNF-α mRNA (potential non-specific effect), 1 nM was used for protein release experiments. Monocyte chemoattractant protein (MCP)-1 (f), IL-6 (g), TNF-α (h), and nitrite (i) release in control (normal) and stably expressing MKP-1 BV-2 cells transfected 24 h with a negative siRNA, or a siRNA against MKP-1 and challenged with LPS (1 μg/ml) for 6 h (MCP-1, IL-6, and TNF-α) or 12 h (nitrite). Results are expressed as mean ± s.e.m. of three experiments in duplicate. *P < 0.05 vs. BV-2 cells stably expressing MKP-1 (pMKP1 +).

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