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. 2010 Dec 9;9:313. doi: 10.1186/1476-4598-9-313

Figure 2.

Figure 2

PEA3 expression and MMP-1 regulation. (A) RT-PCR analysis of PEA3, ER81, MMP-1, MMP-7 and osteopontin mRNA expression in the indicated cell lines. GAPDH was used as a loading control. (B-D) Real time RT-PCR analysis of PEA3, ER81, MMP-1 and the indicated putative target genes in untreated Het1A and OE33 cells (B) or OE33 oesophageal adenocarcinoma cells treated with SMARTpool siRNAs directed against PEA3 (C) or ER81 (D). Average mRNA levels (from duplicate samples in 2 experiments) were calculated relative to the respective mRNA levels of each gene in OE33 cells (B) or to cells treated with a non targeting siRNA (C and D) (taken as 1). (E-G) Real time RT-PCR analysis of PEA3 (E), ER81 (F) and MMP-1 (G) expression in OE33 cells treated with SMARTpool (sp) siRNAs directed against PEA3 or ER81 or one of four individual de-convoluted SMARTpool component siRNAs against PEA3, denoted A-D. Average mRNA expression levels (from duplicate samples in 2 experiments) were calculated relative to the respective mRNA levels of each gene in cells treated with a non targeting siRNA (taken as 1). (H) Reporter gene assay of the activity of a MMP-1-luciferase reporter construct in OE33 cells in the presence of the indicated siRNA duplexes. Cells were co-transfected with either empty vector (grey bars) or with a murine PEA3 expression vector (black bars). Data are shown relative to the activity of the reporter in the presence of non-targeting siRNA duplexes in either the presence or absence of mPEA3 (taken as 1 in both cases) and are the average of 2 experiments performed in duplicate (+/- sem).