Fig. 8: Miconazole effects on activity of NF-κB in LPS-treated mice brain and LPS-treated microglial BV2 cells and cultured astrocytes.
To observe the activity of NF-κB, the expressions of NF-κB-related proteins, such as IκB, p-IκB, p65, and p50 were detected by western blotting using specific antibodies in the mouse brains (a). The activity of NF-κB was measured in 30-m LPS-treated (1 μg/mL) microglial BV2 cells and cultured astrocytes pretreated with MCZ (5 μM and 10 μM) for 1 h. The expressions of proteins, such as IκB, p-IκB, p65, and p50, were detected by western blotting using specific antibodies in the microglial BV2 cells (b) and cultured astrocytes (c). Each blot was representative of three experiments. N: nuclear. The values on the western blot bands represent the arbitrary density measured by ImageJ. The effects of MCZ on LPS-induced NF-κB-dependent luciferase activity in microglial BV2 cells and cultured astrocytes. Microglial BV2 cells (d) and cultured astrocytes (e) were transfected with pNF-κB–Luc plasmid (5 × NF-κB) and then activated with LPS in the absence or presence of MCZ for 12 h, and then, the luciferase activity was determined. Values are the mean and SEM of three independent experiments performed in triplicate. The induction level was calculated relative to the luciferase activity in unstimulated transfected cells. All experiments were performed three times with duplicate. #Significant difference from the LPS-treated group (p < 0.05). *Significant difference from the MCZ-treated group (p < 0.05).
