Anti mouse cd206 mab

Liver tissues were collected and stained with hematoxylin and eosin, and light microscopy was used to observe inflammation and tissue damage. Liver macrophages and neutrophils were detected using primary rat anti-mouse F4/80, CD11b, and Ly6G mAb, respectively (BD Biosciences, San Jose, CA, USA). The secondary, biotinylated goat anti-rat IgG (Vector, Burlingame, CA, USA) was incubated with immunoperoxidase (ABC Kit, Vector), according to the manufacturer’s instruction. Positive cells were counted blindly in 10 HPF/section. iNOS and CD206 in KCs were identified by immunofluorescence using rabbit anti-mouse iNOS mAb and anti-mouse CD206 mAb (Cell Signaling Technology, MA, USA). After incubation with secondary goat anti-mouse Texas Red-conjugated IgG (Sigma, St.Louis, MO, USA), KCs were premounted with VECTASHIELD medium with DAPI (Vector). Negative control slided with the primary antibodies omitted were included in all assays. Positive cells were blindly observed in 10 HPF/section (×200).

LC3B, iNOS, and CD206 in KCs were identified by indirect immunofluorescence staining using rabbit anti-mouse LC3B mAb, anti-mouse iNOS mAb, and anti-mouse CD206 mAb (Cell Signaling Technology, MA, USA). After washing, the KCs which were premounted with VECTASHIELD medium with DAPI (Vector) were incubated with secondary antibody goat anti-mouse Texas Red-conjugated IgG (Sigma, Saint Louis, MO, USA). The numbers of positive-stained cells were counted blindly in 10 high-power field (HPF) sections.

LC3B, iNOS, and CD206 in KCs were identified by immunofluorescence using rabbit anti-mouse LC3B mAb, anti-mouse iNOS mAb, and anti-mouse CD206 mAb (Cell Signaling Technology, MA, USA). After incubation with secondary goat anti-mouse Texas Red-conjugated IgG (Sigma, Saint Louis, MO, USA), the KCs were pre-mounted with VECTASHIELD medium with DAPI (Vector). Positive cells were counted blindly in 10 HPF/section (200×). The positive cells were expressed as a percentage of total cells.