Analysis of glial cell TIMP-2 expression in response to cytokines and LPS. TIMP-2 expression was examined by western blot of cell culture supernatants, as described in Materials and Methods. A. The influence of LPS on TIMP-1 expression in the three different types of glial culture. MGC, astrocytes, and microglial cultures were treated with LPS for 2 days, then cell culture supernatants examined for TIMP-2 expression. Note that MGC and pure astrocytes constitutively expressed TIMP-2 (~22 kD) band, and this expression was not enhanced by LPS stimulation. In contrast, microglial cultures did not express TIMP-2. B. The influence of cytokines on TIMP-2 expression by MGC and pure astrocytes. MGC and pure astrocyte cultures were treated with a panel of different cytokines for 2 days, and the supernatants analyzed for TIMP-1 expression. Note that both MGC and pure astrocytes constitutively expressed TIMP-2, and this expression level was not significantly affected by any of the cytokines tested. C. The influence of serum on microglial TIMP-2 expression. Pure microglial cultures were grown either in serum-free medium or in medium containing 10% FBS. After 2 days, the cell culture supernatants were analyzed for TIMP-2 expression. Note that the presence of serum strongly induced TIMP-2 expression in microglial cells.