Anti tlr2 nb100 56726

The pET30a vector (in our laboratory) was used as an expression vector for N. cyriacigeorgica GUH-2 HBHA in E. coli. The plasmid pK18mobsacB (in our laboratory) was used to construct the hbha deletion mutant. Anti-TLR4 (NB100-56727, Novus Biological, USA), anti-TLR2 (NB100-56726, Novus Biological, USA), and IgG_2A isotype control (MAB003, R&D, USA) were used in the present study. The following antibodies were purchased from Cell Signaling Technology (Danvers, USA): anti-p-Jnk (4668), anti-p-ERK1/2 (4370), anti-p-p38 (4511), anti-p-p65 (3033), and anti-β-actin (7074). The following pharmacological inhibitors were purchased from Sigma-Aldrich (St. Louis, MO, USA): p38 (SB203580), ERK1/2 (PD98059), JNK (SP600125), and NF-κB (BAY 11-7082). The endotoxin removal kit was purchased from GenScript (Nanjing, China). Human and mouse TNF-α, IL-6, and IL-10 ELISA kits were used in this study (BD, USA). DAPI (4′,6-diamidino-2-phenylindole, dihydrochloride) and Alexa Fluor® 488 donkey anti-mouse IgG were purchased from ThermoFisher Scientific (Carlsbad, CA, USA). The adjuvant was purchased from Biodrago (Suzhou, China).

The pET30a vector (constructed in our laboratory) was used to express N. farcinica Nfa34810 in E. coli. The suicide plasmid pk18mobsacB (constructed in our laboratory) was used to construct the nfa34810 deletion mutant. The following antibodies were used in this study: anti-p-Jnk and -JNK (4668; Cell Signaling), anti-p-ERK 1/2 and -ERK 1/2 (4370; Cell Signaling), anti-p-p38 and -p38 (4511; Cell Signaling), anti-p-p65 and -p65 (3033; Cell Signaling), anti-p-AKT and -AKT (4060; Cell Signaling), anti-β-actin (TransGen Biotech), anti-TLR4 (NB100-56727; Novus Biological, USA), anti-TLR2 (NB100-56726; Novus Biological, USA), and IgG_2A isotype control (MAB003; R&D Systems, USA). Human and mouse TNF ELISA kits were used in this study (BD, USA). The following reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA): p38 inhibitor (SB203580), ERK1/2 inhibitor (PD98059), JNK inhibitor (SP600125), and NF-κB inhibitor (BAY 11-7082). An endotoxin removal kit was purchased from GenScript (Nanjing, China).