Sb202190

COX-2 inhibitor NS398 (50 μM, final concentration), p38 inhibitor SB202190 (10 μM), ERK1/2 inhibitor SCH772984 (10 μM), and NF-κB inhibitor JSH-23 (50 μM) (Selleckchem, Houston, USA), as well as ROS inhibitor NAC (10 μM; APEXBIO, Houston, USA) were used to inhibit target proteins in this study. All inhibitors were applied 1 h prior to Giardia exposure.

α‐Minimum essential medium (α‐MEM) and foetal bovine serum (FBS) were obtained from Gibco; Thermo Fisher Scientific, Inc. Soluble recombinant mouse macrophage‐colony stimulating factor (M‐CSF) and recombinant mouse RANKL were obtained from R&D Systems, Inc. Tartrate‐resistant acid phosphatase (TRAP) was obtained from Sigma‐Aldrich and Merck KGaA. High purity Ti particles (average diameter. 1‐5 µm) were obtained from Johnson Matthey (cat. no., 00681). The antibodies (GAPDH, NFkB, C‐FOS, NFATc1, p‐p38,) were purchased from Cell Signaling Technology, Inc. Enzyme‐linked immunosorbent assay (ELISA) kits (IL‐6, IL‐1β, TNF‐α, IL‐10, Arg‐1, iNOS) were purchased from R&D Systems, Inc Flow cytometry anti‐mouse CD16/32‐PE (cat. no.553145) and anti‐mouse CD206‐Alexa 647 (cat. no.565250) were purchased from BioLegend Inc The p38α/β MAPK inhibitor (SB202190) was purchased from Selleck.

COX-2 inhibitor NS398 (50 μM), p38 MAPK inhibitor SB202190 (FHPI) (10 μM), and ERK1/2 MAPK inhibitor SCH772984 (10 μM), was purchased from Selleck Chemicals (Houston, United States). TLR4 inhibitor TAK-242 (10 μM) was purchased from APEXBIO Technology (Houston, United States). Cells were separately pretreated with these drugs at the indicated concentrations for 1 h at 37°C. At the time of the second addition of gentamicin, the inhibitors were also added, and the point was defined as time 0. Cells treated with TAK-242 were collected at 1 h after infection to detect the protein level of phosphorylated and unphosphorylated ERK1/2 MAPK and p38 MAPK, and at 6 h after infection to determine the COX-2 expression level. ERK1/2 inhibitor SCH772984 and p38 MAPK inhibitor SB202190 (FHPI) treated cells were collected at 6 h after infection to detect COX-2 expression level. NS398 treated cells were collected at 6 h and 9 h after infection to detect reactive oxygen species (ROS), cell death, and cytokine expression levels. NS398 treated cells were collected at 6 h after infection to investigate the LC3 expression level. These experiments were performed in triplicate with three biological repetitions.

Rat tail type I collagen, Transwell inserts (0.9 cm² cell culture area, 1.6 × 10⁶ pores/cm²), 12-well polystyrene tissue culture dishes, 12-well polycarbonate plates and Transwell inserts (1.12 cm² cell culture area, 1.6 × 10⁸ pores/cm²) were purchased from Corning (Corning, NY). SB202190 was from Selleckchem (Houston, TX). Collagenase (type IV) was purchased from Worthington Biochemicals (Lakewood, NJ). Y-27632 and N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) were purchased from ApexBio Technology (Houston, TX). Human collagen (type I) was obtained from Advanced Biomatrix (Carlsbad, CA). The ALP assay kit was from AbCam (Cambridge, MA). Gastrin was from Anaspec (Freemont, CA). UGT-Glo™ Assay and P450-Glo™ CYP3A4 Assay were obtained from Promega (Madison, WI). N-acetyl cysteine was purchased from MP Biomedicals (Santa Ana, CA). Murine EGF was procured from Peprotech (Rock Hill, NJ). Primocin was purchased from InvivoGen (San Diego, CA). Nicotinamide, L-alanine p-nitroanilide, fluorescein diacetate, loperamide, zafirlukast, cobalt chloride, potassium phosphate buffer, bovine serum albumin, and A83–01 were acquired from Sigma-Aldrich (St. Louis, MO). All other reagents and Bicinchoninic Acid (BCA) Protein Assay Kit were purchased from Thermo Fisher Scientific (Waltham, MA).