Figure 3
Reduced replication of VSV in oncogene-induced senescent MCF7 cells.
(A) Microscopy images of human tumor MCF7 cells showing morphology (left panels) and SA-beta-gal staining (right panels) of untreated (MCF7-NT, upper panels) and H-Ras-induced senescent MCF7 cells (MCF7-RAS, bottom panels). Quantification of the SA-beta-gal positive cells is shown below (at least 200 cells were counted per condition). (B) Western-blot analysis of H-Ras and downstream targets ERK/P-ERK, and of senescence markers p53 and p21 in untreated (MCF7-NT) or after H-Ras induction (MCF7-RAS) MCF7 cells. GAPDH is shown as loading control. (C) Expression levels of CDKN1A (coding for p21) mRNA relative to GAPDH (x10−3) as determined by qRT-PCR in untreated (MCF7-NT) or H-Ras-induced (MCF7-RAS) MCF7 cells. (D) Western-blot analysis of VSV protein synthesis in untreated (MCF7-NT) or H-Ras-induced (MCF7-RAS) MCF7 cells after the indicated periods of infection at MOIs of 0.05 PFU/cell (left panel) or 10 PFU/cell (right panel). Tubulin is shown as loading control. Data are mean values +/− SE from at least three different experiments. *p < 0.05, **p < 0.01, ***p < 0.001 Student’s t test.
