Figure 5. Increased gelatinase activity in SNAP treated cultures

Cells were cultured for 24 h in serum free media with or without SIN-1 (1 mM or 10 mM) or SNAP (1 mM or 10 mM). The media were harvested, treated with 2 mM of p-aminophenylmercuric acetate (APMA) for one h at 37 °C to activate the MMP-2 from the latent form (72 kDa) to the activated form (68 kDa). A: This is a representative gelatinase zymogram. Lane 1 shows that after activation with APMA (+), the 68 kDa activated MMP-2 is found. Lane 2 shows that in the control, APMA (-) cultures, the major band of gelatinase activity is at 72 kDa that corresponds to the latent form of MMP-2. Lanes 3 and 4 show that in the 10 mM SIN-1 treated cultures, little gelatinase activity is found in either the APMA (+) or APMA (-) samples. Lane 5 shows that the major band in the 10 mM SNAP treated APMA (+) culture is the 68 kDa activated MMP-2. Lane 6 shows that in the 10 mM SNAP treated, APMA (-) cultures, the major gelatinase band is the latent MMP-2 (72 kDa). B: Gelatinase activities found in the APMA (+) activated samples were quantitated by the gelatinase/MMP-2 assay kit. Untreated APMA (+) cultures were normalized to zero. Gelatinase/MMP activities in the 10 mM SIN-1 treated APMA (+) samples were similar to the control APMA (+) samples. Gelatinase/MMP activity in the 1 mM and 10 mM SNAP treated APMA (+) samples were significantly higher compared to control APMA (+) samples (asterisk, p<0.001). Assays were performed in duplicate on different normal cultures (n=5).