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. 2014 Oct 30;13(19):3132–3142. doi: 10.4161/15384101.2014.949212

Figure 4.

Figure 4.

nEGFR and STAT3 cooperatively activated the MMP2 promoter. (A) The sequence near the ATRS in MMP2 promoter is illustrated. The ATRS is underlined. The wild-type and mutant STAT3 binding sites are specified in the open box. The mutated nucleotides in the STAT3 binding motif are capitalized. (B) shControl, shSTAT3-1 and shSTAT3-2 H1975 cells were subjected to RT-qPCR to analyze the MMP2 and STAT3 mRNA expression. (C) 293T cells transfected with pMMP2 Luc, pCMV-NLS-EGFR and the STAT3 shRNA-expression plasmid were subjected to a luciferase reporter gene assay. (D) 293T cells transfected with wild-type or mutant pMMP2 Luc and pCMV-NLS-EGFR were subjected to a luciferase reporter gene assay. (E) 293T cells transfected with pMMP2 Luc, pCMV-NLS-EGFR and/or a STAT3-expressing plasmid were subjected to a luciferase reporter gene assay. (F) Left, H1975 cells were subjected to ChIP with preimmune IgG or an anti-EGFR or anti-STAT3 antibody. Right, H1975 cells were subjected to ChIP with sequential immunoprecipitation with preimmune IgG or an anti-EGFR or anti-STAT3 antibody. The relative amount of the immunoprecipitated MMP2 ATRS DNA fragment was quantified by RT-qPCR with specific primers.