FIG. 2.

DecR1 overexpression partially restores cell contacts in a Neu-transformed breast cancer cell line. (A) Immortalized NMuMG mouse mammary epithelial cells were stably transfected with NeuNT, pooled stable transfectants were injected into the mammary fat pads of nude mice, and tumor cells were explanted back into culture (NMuMG-NT2196). Parental NMuMG, NMuMG/NeuNT-expressing pooled stable transfectants, and NMuMG-NT2196 cells (5 × 10⁴) were injected into the mammary fat pads of nude mice. The data are represented as tumor volumes (mm³) ± standard deviations (n = 4 mice). (B) Generation of NT2196 cells stably expressing DecR1-V5. Whole-cell lysates from NMuMG, NT2196, and three independent NT2196/DecR1-V5 stable cell lines (NT2196/DecR1.1, NT2196/DecR1.7, and NT2196/DecR1.8) were subjected to immunoblot analysis using V5-, Neu-, and α-tubulin-specific antibodies. (C) Quantification of neuNT mRNA levels in NMuMG, NT2196, NT2196/DecR1.1, NT2196/DecR1.7, and NT2196/DecR1.8 cells by real-time PCR. The values were normalized to β2-microglobulin. The values are representative of triplicate samples. (D) DecR1 overexpression partially restores cell contacts in NeuNT-transformed cells. Shown are bright-field images of the indicated cells at confluence (left panels). Cells were also stained with E-cadherin (red) and ZO-1 (green) antibodies and analyzed by confocal microscopy (right panels).