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Copyright by Eunyoung Park 2007 - The University of Texas at Austin

Copyright by Eunyoung Park 2007 - The University of Texas at Austin

FIGURE 3.3. EFFECT OF

FIGURE 3.3. EFFECT OF RETINOL ON MMP MRNA LEVELS. HCT-116 (right column) and SW620 (left column) cells were treated with 0 (ethanol vehicle control), 1, or 10 μM retinol for 24 h. Five different MMP mRNA levels (A: MMP-2, B: MMP-9, C: MMP-7, D: MMP-1 and E: MMP-13) were detected by quantitative real time RT- PCR. Quantitative real time RT-PCR reactions were performed in duplicate. Data are reported as mean ± SEM, n=3. Relative amounts of MMP cDNA were calculated by the comparative CT method. CT values obtained for the different MMP were normalized to corresponding CT values of GAPDH. *Significantly different from control, P < 0.05. 65

FIGURE 3.4. EFFECT OF RETINOL ON MMP ACTIVITY AND PROTEIN LEVELS. HCT-116 (left columns, A and B) and SW620 (right column, C and D) cells were treated with 0 (ethanol vehicle control), 1, or 10 μM retinol for 24 h. Gelatin zymograms (A and C) displaying the active form of MMP-9 (88-92 kDa) and both the pro and active forms of MMP-2 (72 kDa and 62 kDa, respectively) are shown. Western blots (B and D) displaying active (88-92 kDa) MMP-9 protein levels in serum free conditioned media harvested from cells treated for 24 h with retinol. Pro-MMP-9 (95-98 kDa) was not detected in the conditioned media. Each experiment was repeated three times and one representative experiment is shown. 66

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