After rinsing with PBS, sections were incubated with streptavidin-peroxidase conjugate (Dako Corporation, LSAB+ kit, Carpinteria, CA, USA).
Het-1A cells were cultured in Bronchial Epithelial Cell Growth Medium (BEBM) with the additives obtained from Lonza/Clonetics Corporation as a kit CC-31700), without antibiotics.
Serum IGF-1 ELISA (DRG International, USA) and Microvue Osteocalcin EIA (Quidel Corporation, CA, USA) kits were used to measure serum IGF-1 and osteocalcin levels, respectively.
First-strand cDNA was synthesized with a cDNA synthesis kit (Bioneer Corporation, Daejeon, South Korea).
PCR was performed using the GoTaq® Flexi DNA polymerase kit (Promega Corporation, USA).
Immunoreactive bands were detected by an enhanced chemiluminescence detection kit (Millipore Corporation, Billerica, MA, USA) according to the manufacturer's instructions.
Blood samples were collected on the first day of septic shock, and plasma concentration of 23 cytokines/chemokines was assessed using a multi-analyte Milliplex human cytokine kit (Millipore Corporation, Billerica, MA, USA).
DNA was extracted using a DNP™ kit (Cinagen Corporation, Iran).
The sequencing reaction was carried out with the Sequenase Version 2.0 kit (USB corporation).
The assay was performed using OXItek TBARS assay kit (ZeptoMetrix Corporation, Buffalo NY).
Isolation of genomic DNA from W. chondrophila was performed with the Wizard Genomic DNA purification kit (Promega Corporation, Madison, USA).
