Gadolinium chloride hexahydrate (Sigma-Aldrich) powder was reconstituted in sterile saline to a final concentration of 2 mg/mL. To deplete KCs, mice were intraperitoneally given Gadolinium chloride hexahydrate (10 mg/kg) for 2 consecutive days before SAP130 treatment. The same volume of saline was injected as control. To stimulate in vivo Mincle signaling, mice were intraperitoneally injected with SAP130 or TDB (10 mg per mouse). After 2 hours, mice were anesthetized with Zoletil at a dose of 0.2 mL/kg body weight through intramuscular injection before collection of serum and liver tissues for further analysis. 37 C in a 5% CO 2 incubator with RPMI-1640 media that contained 10% fetal bovine serum, 100 IU/mL penicillin, and 100 mg/mL streptomycin. To stimulate Mincle signaling, cells were stimulated with 2 mg/mL recombinant SAP130 (MyBioSource, San Diego, CA) for 24 hours. To inhibit Syk signaling, cells were treated with GS9973 (100 nmol/L) in the presence or absence of recombinant SAP130 for 24 hours.
Primary HPs and KCs were co-cultured in 12-well trans-well plate (Sigma-Aldrich) at a ratio 1:4 of KCs/HPs. To mimic the in vivo condition, cells were cultured with 100 mmol/L EtOH in the presence or absence of LPS (100 ng/mL) for 24 hours.
Primary HPs and KCs were co-cultured in 12-well trans-well plate (Sigma-Aldrich) at a ratio 1:4 of KCs/HPs. To mimic the in vivo condition, cells were cultured with 100 mmol/L EtOH in the presence or absence of LPS (100 ng/mL) for 24 hours.