NADPH oxidase and peroxynitrite–driven hepatic sinusoidal injury via Toll-like receptor (Tlr)-4 signaling. A and B: Representative images of 5-μm-thick immunostained liver sections, imaged using immunofluorescence microscopy. Secondary antibody used was Alexa Fluor 568 (Invitrogen; red) against α-intercellular adhesion molecule (Icam)-1 primary antibody (A) and against α-E-selectin primary antibody (B) (both Icam-1 and E-selectin are sinusoidal injury markers). Nuclear stain (blue) is due to ProLong Gold Antifade Reagent (Life Technologies, Carlsbad, CA) with DAPI. C and D: Morphometry performed on three independent fields for each sample including representative images in for percentage of area showing positive immunoreactivity of Icam-1 in A (C) and of E-selectin in B (D), calculated in arbitrary units. ∗∗P < 0.01. Original magnification, ×20 (A and B). Apo, diet-induced obesity wild-type (DIO) mouse exposed to bromodichloromethane (BDCM) and injected with apocynin; DIO + BDCM, DIO mouse exposed to BDCM; FBA, DIO mouse exposed to BDCM and phenylboronic acid; FeTPPS, DIO mouse exposed to BDCM and iron(III) tetrakis(p-sulfonatophenyl)porphyrin; p47phox KO, high-fat diet–fed mouse with p47phox gene knockout exposed to BDCM; Tlr4 KO, high-fat diet–fed mouse with Tlr4 knockout exposed to BDCM.