Figure 4.

Effect of macrophage-specific $\text{PPAR}\mathrm{\gamma }$ knockout in DEHP-induced fatty liver. Wild-type (WT, $n=5/\text{group}$) and macrophage-specific $\text{PPAR}\mathrm{\gamma }$ knockout (Mac-KO, $n=5/\text{group}$) C57BL/6J male mice were treated with 0.5% (wt/vol) sodium carboxymethylcellulose (CMC; vehicle control) or DEHP ($625\mathrm{mg}/\mathrm{kg}$ BW) by daily gavage for 28 d. (A) Representative image of mice livers and quantification of liver weight and liver index at the end of the treatment ($n=5/\text{group}$). The data are provided in Table S19. (B) Hepatic TG concentration in the liver tissue of WT ($n=5/\text{group}$) and Mac-KO ($n=5/\text{group}$) mice at the end of the treatment. The data are provided in Table S20. (C) Representative images of histology and Oil Red O staining of mouse liver at the end of the treatment (left). Hepatic steatosis and lipid accumulation were quantified by calculating the score of pathological severity and Oil Red O staining intensity, respectively (right, $n=5/\text{group}$). The data are provided in Table S21. (D) Representative flow cytometry gating graph and quantification of hepatic M2 macrophages ($\mathrm{F}4/{80}^{+}\text{CD}{206}^{+}$) in the WT or Mac-KO ($n=5/\text{group}$) mice at the end of the treatment. The data are provided in Table S22. (E) Immunofluorescence staining of CLEC4F (red) and F4/80 (green) in the mouse liver of WT or Mac-KO mice ($n=5/\text{group}$) at the end of the treatment. Cells with positive stain of CLEC4F in the liver sections of WT and Mac-KO mice ($n=5/\text{group}$) were quantified using the ImageXpress system under $20\times \text{magnification}$. The data are provided in Table S23. (F) Representative flow cytometry gating graph and quantification of inflammatory monocyte (${\text{CD}11\mathrm{b}}^{+}{\text{CCR}2}^{+}{\text{Ly}6\mathrm{c}}^{\text{high}}$) and patrolling monocyte (${\text{CD}11\mathrm{b}}^{+}{\text{CCR}2}^{+}{\text{Ly}6\mathrm{c}}^{\text{high}}$) in the liver of WT or Mac-KO mice ($n=5/\text{group}$) at the end of the treatment. The data are provided in Table S24. All data are expressed as $\text{mean}+\text{SD}$. The data were analyzed using two-way ANOVA followed by Tukey’s multiple comparisons test. *$p<0.05$, **$p<0.01$, ***$p<0.001$ compared with the vehicle control (i.e., CMC); significance of comparison between WT and Mac-KO groups of DEHP treatment was labeled above lines. Note: ANOVA, analysis of variance; BW, body weight; DAPI, 4′,6-diamidino-2-phenylindole; DEHP, diethylhexyl phthalate; HE, hematoxylin and eosin; SD, standard deviation; TG, triglyceride.