Non-invasively differentiate non-alcoholic steatohepatitis by visualizing hepatic integrin αvβ3 expression with a targeted molecular imaging modality

Figure 4 Immunofluorescent co-localization of integrin αvβ3 and the markers of various hepatic cells including α-smooth muscle actin, cluster of differentiation 31, cluster of differentiation 68 or cluster of differentiation 163 in livers of the control and high-fat high-carbohydrate diet-fed rats. Representative fluorescent images of integrin β3 subunit (green color) and albumin, α-smooth muscle actin, cluster of differentiation (CD) 31, CD68 and CD163 (red color) in liver sections, which were separately stained with specific first antibodies and visualized by second antibodies, and counterstained with 6-diamidino-2-phenylindole for nuclei staining. The merged images show the yellow color area by overlaying images of the counterstaining, and amplified images corresponding to the indicated areas in boxes are present. Images were recorded at original magnification (400 ×), scale bars = 100 μm. A: Control; B: High-fat high-carbohydrate diet (HFCD)-2M; C: HFCD-4M; D: HFCD-6M rats; E: Comparison of the percentage of integrin β3 subunit positive-staining area in liver sections of the control and HFCD-fed rats. Ten fields were randomly selected and recorded from each section. Then integrin β3 positive-staining area was measured, and the percentages of the positive-staining area in liver sections were compared; F: Comparison of the ratio of the overlapped yellow area to integrin β3 subunit positive-staining green area in liver sections of HFCD-6M rats. Ten fields were randomly selected and recorded from each section. Then integrin β3 subunit positive-staining area and the area of integrin β3 subunit positive-staining overlapped with hepatic cellular markers positive-staining were respectively measured, and the ratios were compared. In all panels, data are expressed in means ± SD. DAPI: 6-diamidino-2-phenylindole; CTRL: Normal control group; M: Month; CD: Cluster of differentiation; αSMA: α-smooth muscle actin.