Formalin-fixed, paraffin-embedded 5 μm intestinal slices from Lean Control, (Chow+PP2Ainh), NAFLD, (NAFLD+PP2Ainh) groups were used for immunofluorescence imaging. (A) (i-iv) Immunofluorescence images depicting gp91phox (red) and p47phox (green) co-localization events in the small intestine, counterstained with DAPI (blue) of Lean Control, (Chow +PP2Ainh), NAFLD and (NAFLD+PP2Ainh) mice groups. Images were taken at 40X magnification, (B)(i-iv) Immunofluorescence images depicting the proximity ligation (red) of gp91phox and p47phox proteins in the small intestine, counterstained with DAPI (blue) of Lean Control, (Chow+PP2Ainh), NAFLD, (NAFLD+PP2Ainh) mice groups. Images were taken at 40X magnification. (C) Morphometric analysis of gp91phox- p47phox co-localization events. Y-axis shows % positive immunoreactive area (% ROI) (n=3, analysis from three separate microscopic fields) (0.05<**p < 0.01). (D) Morphometric analysis of gp91phox and p47phox proximity ligation. Y-axis shows % positive immunoreactive area (%ROI) (n=3, analysis from three separate microscopic fields) (#p < 0.01). (E and F) Correlation analysis of fecal and serum lactate with NOX2 activation. Significance was tested by performing unpaired t-test between the groups (*p< 0.05, 0.05<**p < 0.01, #p< 0.01), followed by Bonferroni Dunn Post hoc corrections. Results were expressed as mean ± SEM.