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Figure 2. Expression of MMP-2, MMP-9, and VEGF-A in primary AML cells. (A) PCR analyses of MMP-2/9 and VEGF-A transcripts. Samples were standardized for total cDNA content by assessing the presence of identical amounts of β2-microglobulin transcripts. (B) Expression profiles of MMP-2/9 (pro and active forms) were analyzed by zymography using 7.5% (w/v) SDS-polyacrylamide gels containing 0.1% gelatin (w/v) in the 72-h conditioned media (supernatant) from AML cells. The control medium was an FCS-supplemented culture medium alone incubated under the same conditions; the culture supernatant from U937 (M5) cells was used as a positive control for proMMP-2/9 and active MMP-9 proteins. The assay’s sensitivity for MMP-2/9 gelatinolytic activity was 25 ng/mL. (C) Total MMP-2 (first column, n = 27 samples), total MMP-9 (second column, n = 27 samples), and VEGF-A (third column, n = 17 samples) productions in the 72-h culture supernatants from AML cells were determined by ELISA. Mean concentrations ± SEM are indicated. Spots are superposed in all columns. (D) Correlations among MMP-2, MMP-9, and VEGF-A levels in the 72-h culture supernatants from AML cells. Spearman’s correlation coefficient (r) and the P-value are shown for MMP-2 vs. MMP-9 (n = 27), MMP-2 vs. VEGF-A (n = 17), and MMP-9 vs. VEGF-A (n = 17).