Anti f4 80

Hematoxylin and eosin (H&E) staining and immunostaining of tissues were performed according to standard protocols on 5-μm-thick cryostat sections. For immunostaining, primary antibodies included anti-MMP-12 (sc-30072, Santa Cruz Biotechnology, Dallas, TX), anti-CD68 (AbD serotec, Raleigh, NC), anti-F4/80 (Cedarlane, Burlington, NC) and anti-CD31 (BD Pharmingen, San Jose, CA). Isotype-matched antibodies were used as controls. For co-immunostaining, tissue sections were fixed with acetone, and stained with the antibody followed by probe 2. Nuclei were stained with DAPI using ProLong^® Gold Antifade Mountant with DAPI (Molecular Probes, Eugene, OR). The slides were photographed using a Spot RT3 camera (Diagnostic Instruments, Sterling Heights, MI) and a Zeiss Axiophot fluorescence microscope (Carl Zeiss Microscopy GmbH, Jena, Germany).

Portions of the liver were excised and fixed immediately with 4% formaldehyde at room temperature. Paraffin-embedded tissue sections were cut into 4 μm slices and placed on slides. Sections were stained with hematoxylin and eosin or Sirius red, according to standard procedures. Anti-F4/80, anti-CD11c, and anti-Ly6C antibodies were purchased from Cedarlane Laboratories (Burlington, Ontario, Canada), Invitrogen (Waltham, MA, USA), and Abcam (Cambridge, MA, USA), respectively. Positive areas for F4/80 and Sirius red were measured using ImageJ software Version 1.53t [45 (link)]. Histologic steatosis, lobular inflammation, and hepatocyte ballooning were assessed according to the criteria proposed by Kleiner et al. [46 (link)]. All histological analyses were performed by pathologists (K.Tsuneyama and M.I-S.), and the histological scores and grade were determined in a blinded manner.

Portions of the liver were excised and fixed immediately with 4% formaldehyde at room temperature. Paraffin-embedded tissue sections were cut into 4-μm slices and placed on slides. Sections were stained with hematoxylin and eosin or Sirius red, according to standard procedures. Anti-F4/80, anti-CD11c, and anti-Ly6C antibodies were purchased from Cedarlane Laboratories (Ontario, Canada), Invitrogen, and Abcam (Cambridge, MA, USA), respectively. Positive areas for F4/80 and Sirius red were measured using ImageJ software [62 (link)]. Histologic steatosis, lobular inflammation, and hepatocyte ballooning were assessed according to the criteria proposed by Kleiner et al. [11 (link)]. All histological analyses were performed by pathologists (K.Tsuneyama and M.I.-S.), and the histological scores and grade were determined in a blinded manner.