Figure 1

Ras ^G12V induces CXCL8 expression independently of deregulated p53, and synergizes with the inflammatory cytokines TNFα and IL-1β. MCF-7 cells were transfected to express p53^shRNA, Ras^G12V, Ras^G12V + p53^shRNA or the appropriate control vectors. (A, B) Induction of CXCL8 by Ras^G12V ± p53^shRNA expression, determined in cell CM at the protein level by ELISA (A), or at the mRNA level by qRT-PCR (B). (C, D) Induction of CXCL8 expression by the synergistic activities of Ras^G12V with IL-1β (500 pg/ml) or TNFα (50 ng/ml), determined at the protein level by ELISA (C), and at the mRNA level by qRT-PCR (D). Cytokine concentrations were selected based on previous titration analyses. *p < 0.05, **p < 0.01, ***p < 0.001 compared to control transfectants (A, B), or to non-stimulated cells (C, D). NS = Not significant. In all panels, a representative experiment of n≥3 is presented. Please see “Methods” for additional details on times of CM collection, and of mRNA analyses.