Figure 7
The NG-to-HG shift enhances LPS-induced inflammation in cultured BV-2 microglia. (a) As indicated, both NG- and HG-cultured cells were simply treated with or without LPS (1 µg/ml); also, an NG-to-HG shift with or without LPS treatment in NG-cultured cells was performed. After incubation for 24 h, western blotting was used to detect iNOS and COX-2 levels (for uncropped images of the blots please see Supplementary Information). GAPDH served as a protein loading control. After normalization to corresponding GAPDH, iNOS and COX-2 levels were quantified using densitometry and expressed relative to the constant NG group (assigned a value of 1). The mean ± SEM from four independent experiments is shown below the representative blots. *p < 0.05 versus constant NG group treated with LPS. (b–e) Both NG- and HG-cultured cells as well as the NG-cultured cells receiving an NG-to-HG shift were incubated with different concentrations of LPS (0.001–10 µg/ml; b) or single LPS concentration (1 µg/ml; c–e) for 24 h. ARA-C was added to cultures to avoid the influence of cell proliferation rates. After harvest, cells were subjected to various assays for determining nitrite accumulation (b), TNF-α release (c), and productions of peroxides (d) and ROS (e). Data are represented as mean ± SEM from four to five independent experiments performed in triplicate. *p < 0.05 versus constant NG group at equal LPS concentration (b) or constant NG group treated with LPS (c–e); #p < 0.05 versus NG-to-HG group treated with LPS.
