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. 2012 Mar 26;287(21):17109–17119. doi: 10.1074/jbc.M111.326975

FIGURE 4.

FIGURE 4.

Effects of altered S100A14 expression on MMP2 expression and the promoter activity of MMP2 are dependent on p53 status. A, H1299, HT1080, and MCF-7 cells were transfected with the indicated constructs, and pooled neomycin (G418)-resistant colonies were established as stable transfectants. The protein expression of S100A14, p53, and MMP2 was determined by Western blot, and β-actin served as a loading control (left panel); the mRNA expression of MMP2 was determined by RT-qPCR (middle panel); and MMP2 transcriptional activity was detected by MMP2-luc reporter assay (right panel). B, siRNAs targeting S100A14 (25 nm) (ON-TARGETplus SMARTpool) or control siRNAs were transfected into HCT116/p53+/+ and HCT116/p53−/− cells, and 72 h later, proteins and mRNA were extracted and subjected to Western blot or RT-qPCR. C, cell invasion assay was performed in S100A14-overexpressed H1299, HT1080, and MCF7 cells. Representative pictures (left panel) and relative numbers of invading cells (right panel) are shown. Cells were counted in four randomly selected fields. Error bars represent the S.E. of triplicate experiments. *, p < 0.05; **, p < 0.01 two-tailed Student's t test. D, stable-silenced S100A14 HCT116 cells were obtained, and proteins and RNA were extracted and subjected to Western blot or RT-qPCR (left panel). pGC-1, control shRNA-transfected clone; S1-7 and S2-8, shRNA targeting S100A14-transfected clones. Cell invasion assay was performed in stable-silenced S100A14 HCT116 cells (right panel).