PML repressed nEGFR-induced MMP2 expression. (A) RT-qPCR was performed to determine the relative mRNA expression of PML, MMP2 and MMP9 in H1975-shControl, H1975-shPML and H1975-shPML-2 cells as described in the Materials and Methods. (B) H1975 shPML cells with or without overexpression of shRNA-resistant PML (rPML) were analyzed by RT-qPCR to determine the relative mRNA expression of PML and MMP2. (C) 293T cells transfected with pMMP2 Luc and 100 ng of pCMV-EGFR NLS or pCMV-EGFR were analyzed using the luciferase reporter gene assay as described in the Materials and Methods. (D) A549 cells were transfected with pMMP2 Luc and increasing amounts of the pHA-PML expression plasmid. Twenty-four hours after transfection, the cells were treated with or without 50 ng/ml EGF for 18 h and then subjected to a luciferase reporter gene assay. (E) 293T cells were transfected with pMMP2 Luc, 100 ng of pCMV-EGFR NLS or empty vector and increasing amounts of PML expression plasmid as indicated. The cells were subjected to a luciferase reporter gene assay 48 h after transfection. (F) 293T cells were transfected in 6-well plate with 1 μg of pCMV-EGFR NLS or empty vector, and increasing amounts of pHA-PML as indicated. Forty-eight hours after transfection, the cells were analyzed by RT-qPCR to detect MMP2 mRNA expression.