Figure 4. Effect of ablation of IκBα on MMP-2 expression and activity 24 h, 48 h, and 72 h after IκBα siRNA transfection. A:MMP-2 mRNA expression in in HCM cells of IκBα siRNA transfected, nonsense control siRNA (NC) transfected, and control was quantified by real-time RT-PCR. Expression levels were normalized with GAPDH. B: After IκBα siRNA transfection, the conditioned media were collected at the indicated time, concentrated, and analyzed by western blot with MMP-2 antibody. The arrow indicates MMP-2 bands that are analyzed by densitometry and the values were represented in the bar graph. C: The activity of MMP-2 is analyzed by gelatin zymography analysis. The arrows indicate pro-MMP-2 (72-kDa) and active MMP-2 (66-kDa) specific bands. The bands were analyzed by densitometry and are represented in the bar graph. The mRNA, protein, and activity values of IκBα siRNA transfected, nonsense control siRNA (NC) transfected, and control were determined by one-way ANOVA. An asterisk denotes p<0.05, and a double asterisk indicates p<0.01.