Lipopolysaccharide lps from e coli o127 b8

Dendritic cells (BM-DCs) were generated from the bone marrow extracted from femur and tibia of 7–10-week-old mice as described previously (Quast et al., 2011 (link)). We employed DCs from Lifeact-EGFP mice, which allow visualization of F-actin without interfering with polymerization dynamics (Riedl et al., 2010 (link)). In brief, bone marrow cells were collected by flushing the bones with PBS (without Ca²⁺, Mg²⁺; Pan Biotech). Subsequently, 5 × 10⁶ bone marrow cells were cultured in 10 cm petri dishes (Greiner Bio-one) in 10 ml complete cell culture medium [VLE-RPMI 1640 (Pan Biotech), supplemented with 10% Fetal Calf Serum (FCS, Sigma-Aldrich, Pan Biotech), 100 µ/ml Penicillin (Pan Biotech), and 100 μg/ml Streptomycin (Pan Biotech)] containing 10 ng/ml recombinant Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF, Peprotech). On day 3, cells were fed by addition of 5 ml complete cell culture medium supplemented with 10 ng/ml GM-CSF. On day 6, half of the cell culture medium in stock was replaced by fresh complete cell culture medium supplemented with 10 ng/ml GM-CSF. To induce maturation, BM-DCs were stimulated overnight with 200 ng/ml Lipopolysaccharide (LPS) from E. coli O127:B8 (Sigma-Aldrich) and used for experiments on day 7–9.

Sitagliptin (SIT; cat. No. HY-13749) and caerulein (HY-A0190) were obtained from MedChemExpress (Princeton, NJ, USA). Lipopolysaccharide (LPS; from E. coli O127: B8) was purchased from Sigma (St. Louis, MO, USA). Antibodies against LC3-ll (4108), Beclin1 (3738), Atg5 (2630), and NQO1 (62262) were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA); MPO (ab9535), 4-HNE (ab46545), DPP4 (ab187048), and Lamin B1(ab16048) were purchased from Abcam Inc. (Cambridge, MA, USA); and β-actin (66009-1-Ig), Nrf2 (16396-1-AP), Keap1 (10503-2-AP), Ho-1 (10701-1-AP), and p62 (18420-1-AP) were purchased from Proteintech Inc. (Shanghai, China).

Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), and trypsin were purchased from Gibco (Grand Island, NY). Sitagliptin (sit; cat. No. HY-13749) was purchased from MedChemExpress. Lipopolysaccharide (LPS; from E. coli O127 : B8) was from Sigma. Cerulein (HY-A0190) was from MedChemExpress. Antibodies to DPP4 (ab187048), IL-6 (ab6672), IL-1β (ab9722), and TNF-α (ab6671) were from Abcam Inc. (Cambridge, USA). Antibodies to GAPDH (5174S), Nrf2 (12721), and NF-κB (8242) were from Cell Signaling Technology Inc. (Beverly, MA, USA).