C57BL/6NCrlBR male mice (8–10 weeks, Charles River) were challenged by intratracheal (i.t.) administration with an average of 5.6 × 105 colony forming units (CFUs) of the wt and ∆lasB RP45 strains or 3 × 105 CFUs of the wt and ∆lasB RP73 strains embedded in agar-beads (Bragonzi et al., 2009; (link)
Facchini et al., 2014 (link); Cigana et al., 2016b (link)) or with sterile empty agar-beads as control. Body weight and health status were monitored daily. Lung CFUs were analyzed at Days 2 and 7 post-infection. As reported previously, recovery of ≥1,000 CFUs of P. aeruginosa from Day 7 lung cultures is considered evidence that a chronic infection has been established (Lore et al., 2016 (link), 2018 (link)). Cytokine/chemokine/growth factor levels were measured in the supernatant of lung homogenates by Bio-Plex Assay (Bio-Rad), as described in the online data supplement. Additional details in accordance with the ARRIVE guidelines (Kilkenny et al., 2010 (link)) are reported in the online data supplement.
Facchini et al., 2014 (link); Cigana et al., 2016b (link)) or with sterile empty agar-beads as control. Body weight and health status were monitored daily. Lung CFUs were analyzed at Days 2 and 7 post-infection. As reported previously, recovery of ≥1,000 CFUs of P. aeruginosa from Day 7 lung cultures is considered evidence that a chronic infection has been established (Lore et al., 2016 (link), 2018 (link)). Cytokine/chemokine/growth factor levels were measured in the supernatant of lung homogenates by Bio-Plex Assay (Bio-Rad), as described in the online data supplement. Additional details in accordance with the ARRIVE guidelines (Kilkenny et al., 2010 (link)) are reported in the online data supplement.
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