Effects of NNMT overexpression on the expression of MMP-2. (a) The expression levels of MMP-2 mRNA in HEK293/vector (control) and HEK293/NNMT cells were determined by Q-PCR and normalized to GAPDH. Data are presented as the mean of triplicate replications. S.D. is indicated by error bars. ** versus control cells. (b) Gelatin zymography assay was used to examine the MMP-2 activity in conditioned media, and western blot analysis to detect MMP-2 and NNMT in cell lysates from HEK293 cells with or without NNMT expression. (c) HEK293/vector and HEK293/NNMT cells were transfected with 40 μmol/L of NNMT siRNA or control siRNA. Q-PCR (upper) gelatin zymography and Western blot analyses (lower) were performed to determine the expression levels of NNMT and MMP-2. Q-PCR data are presented as the mean of triplicate replications. S.D. is indicated by error bars. ** versus control cells. Actin was used as loading control.
