Figure 3

STAT3 activation and cytokine production in cultured cells. A, Phospho-Tyr705-STAT3 and total STAT3 were immunoblotted after treatment with LPS (100 ng/mL) for 6 h of primary cultures of astrocytes, microglia, or neurons. Some of the neuronal cultures also were treated for 6 h with conditioned media (CM) from LPS-treated (6 h) astrocytes. B, Primary enriched astrocytes were treated with vehicle (0), or with 100 ng/mL LPS, 1 ng/mL IFNγ, or both, for 2, 4, 6, or 8 h, followed by measurements of IL-6. Means ± SEM; n = 5; *p < 0.05 compared to LPS alone. C, Primary enriched astrocytes were stimulated with LPS (100 ng/mL) and IFNγ (1 ng/mL), with or without the selective GSK3 inhibitor lithium (LiCl, 20 mM), for 6 h. Culture supernatants were incubated with an array membrane that detects 62 proteins in duplicate. Boxes indicate the location of the detected 13 proteins induced by LPS plus IFNγ. Positive controls are in the upper left and lower right corners (high intensity spots) and negative controls in the upper right corner (no spots).