Mueller hinton broth

The PAO1 strain (ATCC 15692) used in this study was kindly provided by Mingqiang Qiao (College of Life Sciences of Nankai University, Tianjin, P.R. China). P. aeruginosa PA47 is a clinical isolate and strong biofilm producer, as we previously reported (She et al., 2018). Other clinical isolates of P. aeruginosa were collected from patients with pulmonary infections at the Third Xiangya Hospital of Central South University (Changsha, Hunan, P.R. China) from January 2014 to December 2014 (Qu et al., 2016). Detailed information on other strains and plasmids used in this study is shown in Table 1. All strains were stored at −80°C in whole milk culture. Glucose‐free Luria‐Bertani (LB) broth (Solarbio) was used for bacterial planktonic cell growth and biofilm formation in all experiments; Mueller‐Hinton (MH) broth (Solarbio) was used for the antibiotic susceptibility assay.

Imipenem was purchased from Aladdin Technology (China). Fetal bovine serum (FBS) and high glucose (Dulbecco’s modified Eagle’s medium DMEM) were obtained from Life Technologies. Dialysis membranes (2000 D), hematoxylin and eosin (HE), crystal violet (CV), and phenazine methosulfate were provided by Solarbio Technology (China). Mueller-Hinton (MH) broth was obtained from Solarbio. Polycarbonate porous membrane syringe filters (200 nm) were obtained from Whatman. The bacterial stock solution and reactive oxygen species (ROS) assay kits were purchased from Beyotime Technology (China). The LIVE/DEAD backlight bacterial viability kit was purchased from Yeasen Biotechnology (China). Anti-IL-6 and anti-Ly6G antibodies were manufactured by BOSTER Biological Technology (China). The HRP-conjugated goat anti-mouse IgG was purchased from BOSTER Biological Technology.