Figure 11.

Effects of LY-294002 on MMP-1 (A), MMP-2 (B), and MMP-3 (C) secretion from ONH astrocytes at 6 hours. ONH astrocytes were starved in serum-free medium for 16 hours. Cells were then pretreated with LY-294002 (1 μM) for 30 minutes followed by SNC-121 (1 μM) and TNF-α (25 ng/mL) treatments for 6 hours. Secreted media was collected and concentrated 10-fold. An equal volume (40 μL) of conditioned media was analyzed by Western blotting using selective anti–MMP-1, anti–MMP-2, and anti–MMP-3 antibodies followed by incubation with appropriate secondary antibodies (HRP-conjugated; dilution 1:3000). The signal was captured using enhanced chemiluminescent reagent and the Biorad Versadoc imaging system. Data are expressed as mean ± SE. *P < 0.05; n = 4–7.