Figure 3

InvColi-pS strains infect CRC cells and induce high COX-2 silencing. GFP protein expression was evaluated in (A) HT-29 and in (B) HCA-7, 48 h after InvColi- pS^TBE infection (bar=30 μm). (C) PCR analysis of Inv, HlyA and pTBE loci was performed on pGB2-Ω-inv-hly- and pS^TBE-purified plasmids, on lysed InvColi-pS^TBE bacteria and on InvColi-pS^TBE-infected HT-29 cells. PCR products were analysed after electrophoresis on 2% agarose gel. COX-2 protein, COX-2 mRNA and siCOX-2 levels were detected in (D) HT-29 and (E) HCA-7 cells, 48 h after InvColi strain infection, by western blot and real-time PCR. COX-2 protein, COX-2 mRNA and siCOX-2 expression was normalised against β-actin protein, GUSB (β-glucuronidase) mRNA and U6 RNA levels, respectively. Relative expression of COX-2 protein, COX-2 mRNA and siCOX-2 refers to InvColi-pS^H1-infected sample. Data represent mean±s.e.m. of three independent experiments. ^*P<0.01.