Figure 1

Schematic representation of Giant Plasma Membrane Vesicles production and purification. Briefly, the cells were treated with GPMV-reagent containing PFA and DTT during 1 h to allow vesiculation. The starting cell culture should be prepared in a culture area 8 × higher than the surface where the purified GPMVs will be applied, in order to achieve a confluent layer of vesicles. After incubation, GPMV-enriched supernatant was collected and centrifuged to sediment cellular debris. The clear GPMVs suspension was dialyzed to remove the PFA and DTT. The GPMV suspension was also concentrated and seeded in a poly-L-lysine coated plate, overnight at 37 °C, 5% CO₂ for further assays. For adhesion assays, 24-well plates were used, and adhesion was performed with 5 × 10⁷ B. cenocepacia K56-2 CFU per well. For total RNA extraction and further RNA sequencing analysis, adhesion was performed in a culture plate of Ø 100 mm using 1.5 × 10⁹ B. cenocepacia K56-2 CFU per plate. In both cases, bacteria were allowed to adhere for 30 min, at 37 °C, 5% CO₂. The figure was created using materials from SMART- Servier medical art (https://smart.servier.com/).