FIGURE 3:

SRF mediates induction of EMyT markers in γ-CYA–depleted epithelial cells. (A, B) A549 cells transfected with either control or γ-CYA siRNAs were immunolabeled for SRF and counterstained with a nuclear dye, DAPI, on day 4 posttransfection. Intensity of nuclear SRF signal was quantified with image analysis. (C) A549 cells stably expressing SRE luciferase reporter were transfected with either control or γ-CYA siRNAs and cotransfected with a Renilla luciferase plasmid. Dual luciferase assay was performed 2 d posttransfection. The luminescence signal of SRF-dependent promoters was normalized by the Renilla luciferase signal. (D, E) A549 cells were subjected to sequential transfections with one of the following siRNA pairs: control–control, SRF–control, control–γ-CYA, and SRF–γ-CYA. Expression of targeted proteins and EMyT markers was determined by immunoblotting on day 3 after the second transfection. Data are presented as mean ± SE (n = 3); **p < 0.005, ***p < 0.0005.