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Figure 10 | Journal of Neuroinflammation

Figure 10

From: Glycogen synthase kinase-3β inactivation inhibits tumor necrosis factor-α production in microglia by modulating nuclear factor κB and MLK3/JNK signaling cascades

Figure 10

GSK-3β-mediated NF-κB and MLK3/JNK are two independent signaling pathways in the induction of TNF-α. (A) BV-2 cells were pretreated with SP600125 (SP, 10 μM), K252a (50 nM), BAY 11-7082 (BAY, 5 μM) or PDTC (20 μM) for 30 min following exposure to 100 ng/ml LPS for another 30 min. IκB-α degradation was determined by western blotting. (B) NF-κB luciferase reporter assays were performed by LPS stimulation in the absence or presence of 10 μM SP600125, 50 nM K252a or 20 μM PDTC (6 h) of BV-2 cells as described in Fig. 3. Data are presented as mean + SEM for three independent experiments. **p < 0.01 compared with LPS alone. (C) Cells were treated as described in (A). Phosphorylated and total JNK and c-Jun were detected by western blotting in whole cell extracts. (D) Cells were preincubated with SP600125 (2.5 μM), K252a (12.5 nM), BAY 11-7082 (0.625 μM), SP600125 plus BAY 11-7082 or K252a plus BAY 11-7082 for 30 min, and then treated with 100 ng/ml LPS for 6 h. Released TNF-α was measured by ELISA. Data are presented as mean ± SEM for three independent experiments. #p, ##p or *p < 0.01 compared with SP600125, K252a or BAY 11-7082 treated cells, respectively. The TNF-α content in untreated cultures was not detectable. The level of TNF-α in cells treated with LPS alone was 5.74 ± 0.41 ng/ml.

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