P-FPMI-224 - P-FPMI-224
Mid-exponential phase broth cultures of bacteria (Pseudomonas aeruginosa) were added to the apical surface
of polarized cells at an MOI of 50 viable bacteria per cell.
16HBE14O- epithelial cells and bacteria were incubated together for 1h in complete MEM.
The supernatants containing the bacteria were then replaced by fresh sterile medium and cells were incubated for a
further 3 hours, after which they were lysed with the RLT lysis buffer supplemented with β-mercaptoethanol
(RNeasy Mini kit, with RNase-Free DNase treatment (Qiagen, Hilden, Germany)).