Effects of colibactin on interleukin 8 secretion and nuclear factor κB translocation in human hepatic epithelial cells.

Mock-KO, ATG5-KO and ATG7-KO Hep3B cells were infected for 3 days with colibactin-secreting extra-intestinal pathogenic E. coli (pks⁺) and its corresponding isogenic mutant (pks^-). (A) IL8 secretion was then quantified on cell culture supernatants (Human IL-8/CXCL8 Quantikine ELISA Kit, R&D Systems, Minneapolis, MN, USA) and (B) cells were processed for fluorescent staining with primary antibodies generated against the p65 subunit of nuclear factor κB (NF-κB) and DAPI to counterstain the nuclei (blue). Nuclear translocation of the NF-κB was quantified on a minimum of 500 cells using the "Integrated Density" measure function of ImageJ. *p< 0.005, **p< 0.001, ***p< 0.0001. Abbreviations: AU, arbitrary unit; KO, knock-out; ns, non-significant; pks^-, bacterial artificial chromosome vector; pks⁺, bacterial artificial chromosome vector with pks island encoding colibactin.

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