Control igg

A total of 2 × 10⁵ PBMCs from patients with CHB were cultured in DMEM (HyClone SH30022.01) supplemented with 10% FBS and IL-2 (100 IU/ml), in the presence of an anti-human NKG2A blocking antibody (CloneZ199, Beckman Coulter, United States) or control IgG (BD Biosciences) at 37°C in 24-well plates. After 7 days, the phenotype and function of NK cells were analyzed by flow cytometry.
PBMCs (2 × 10⁵) isolated from healthy donors were seeded into 24-well plates in DMEM in 20% serum from healthy controls containing 100 IU/ml IL-2, then 500 ng/ml HBeAg (Prospec, HBV272) was added into the wells and cells were cultured for 7 days at 37°C. In the presence of HBeAg, 50 ng/ml anti-human IL-10 neutralizing antibody (Clone25209, R&D, United States) or control IgG (BD Biosciences) was added. The intracellular cytokine in NK cells and the cytokine secreted in the supernatant were measured by flow cytometry.

Cells treated for 2 hours with eribulin or vehicle were lysed in a CHAPS buffer containing protease and phosphatase inhibitors. Equivalent amounts of protein were precleared with protein G beads (GE Healthcare), immunocomplexes containing p130Cas-IgG or control-IgG (BD Biosciences) were formed for 2 hours and then protein G beads used to precipitate the immunocomplexes. Beads were washed and prepared for immunoblotting.

We induced dermal fibrosis in mice by bleomycin as previously described [29 (link)]. Bleomycin was dissolved in saline at 1 mg/ml. The saline or bleomycin was administered subcutaneously into the shaved back of the mice (male 8-week-old SCID mice). The administration of saline or bleomycin in the same site was carried out daily for up to 2 weeks. In other experiments, the saline, bleomycin, saline plus clodronate (FormuMax Scientific, CA, USA), or bleomycin plus clodronate were similarly administered to the mice (male 8-week-old C57BL/6 J mice). The administration of saline or bleomycin in the same site was carried out daily for up to 2 weeks, and the administration of clodronate was carried out intraperitoneally every 3 days for up to 2 weeks. In addition, the saline or bleomycin was similarly administered to the mice (male 8-week-old C57BL/6 J mice). The administration of saline or bleomycin in the same site was carried out daily for up to 2 weeks, and the administration of control IgG (2 mg/kg) or anti-IL-4Rα antibodies (2 mg/kg) (BD Biosciences, NJ, USA) was administered intraperitoneally every week for up to 2 weeks.