F4/80 Antibody - #DF2789

Figure 1 LPS/D-GalN-induced ALF enhances apoptosis and ferroptosis. (A) The survival rate of WT mice treated with vehicle or LPS/D-GalN (LPS, 20 μg/kg; D-GalN, 700 mg/kg) (n = 10). (B) Representative pictures of WT mice treated with vehicle or LPS/D-GalN for 3, 5, and 6 hours (n = 10). (C) Serum levels of ALT and AST with or without LPS/D-GalN co-injection (n = 6). (D) H&E and TUNEL staining after LPS/D-GalN co-injection (n = 6). (E) Immunohistochemistry of TNFα and F4/80 staining after LPS/D-GalN co-injection (n = 6). (F) Transmission electron microscope of liver tissues (n = 3). (G) MDA assay of liver homogenates (n = 5). (H and J) Western blot analyses of TFR, DMT1, GPX4, and XCT from WT mice (H) with or without LPS/D-GalN and (I) quantitative results (n = 3–9). (I and K) FSP1, DHODH, and POR proteins. Western blot analyses of WT mice (I) with or without LPS/D-GalN and (K) quantitative results (n = 3–9). (L) Immunohistochemistry of DMT1 staining and immunofluorescence of Ptgs2 in livers with or without LPS/D-GalN (n = 6). (M) Quantitative results of DMT1 and Ptgs2. All data were obtained from WT mice. Scale bars: 100 μm. Data are presented as means ± SEM. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. DAPI, 4′,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Fig. 6. |Pharmacologic Inhibition of iNOS Function Attenuates DSS Colitis. (a) Loss of body weight was recorded from day 0 to day 7. (b) Colon lengths were described and quantified in a bar graph. (c) Serum NO level was quantified in a bar graph. Immunohistochemistry analysis was utilized for the F4/80 (d) and Claudin-1 (e).

Fig. 2. |Down-regulation of Cav-1 in the colon tissue and accompany with the increase of INOS and NO levels after DSS administration. (a) Western blot analysis for detecting Cav-1, Bcl-2, P-Stat3 and Stat3 expression incolon tissues and quantified in a bar graph. (b) Immunofluorescence analysis for detecting Cav-1 and the macrophage-specific marker F4/80 expression in the colon tissue

Fig. 5 CX3CL1 deficiency attenuated kidney inflammation and macrophage infiltration in cisplatin-induced model mice. A Western blotting was conducted to assess the expression profiles of IL-6, TNF-α, and F4/80 in kidney tissues. B, C Levels of IL-6 and TNF-α in mouse sera. D Image representations of TNF-α and F4/80 expression by immunohistochemistry. Scale bar = 20 μm. E podocytes were transfected with siRNA-CX3CL1 as indicated and treated with cisplatin (10 μM) for 24 h. CX3CL1, TNF-α, and IL-6 expression profiles were then measured by western blotting. F, G Levels of IL-6 and TNF-α in podocyte cell supernatant. H, I Representative micrographs of CX3CL1 and TNF-α staining in podocytes. Scale bar = 10 μm. (IL-6, interleukin 6; TNF-α, tumor necrosis factor-α; P value was calculated by one-way analysis of variance and Tukey’s test.

Fig. 5 CX3CL1 deficiency attenuated kidney inflammation and macrophage infiltration in cisplatin-induced model mice. A Western blotting was conducted to assess the expression profiles of IL-6, TNF-α, and F4/80 in kidney tissues. B, C Levels of IL-6 and TNF-α in mouse sera. D Image representations of TNF-α and F4/80 expression by immunohistochemistry. Scale bar = 20 μm. E podocytes were transfected with siRNA-CX3CL1 as indicated and treated with cisplatin (10 μM) for 24 h. CX3CL1, TNF-α, and IL-6 expression profiles were then measured by western blotting. F, G Levels of IL-6 and TNF-α in podocyte cell supernatant. H, I Representative micrographs of CX3CL1 and TNF-α staining in podocytes. Scale bar = 10 μm. (IL-6, interleukin 6; TNF-α, tumor necrosis factor-α; P value was calculated by one-way analysis of variance and Tukey’s test.

FIGURE 4 VTE inhibits hepatic inflammation in MCDD-fed mice. Male C57BL/6J mice (20 ± 2 g) were fed an MCD diet in the presence and absence of VTE (1 g 100 g−1 diet) for 6 weeks (A) F4/80 immunofluorescence analysis in the livers from the MCS diet-, MCD diet- and MCD + VTE-fed mice. Scale bars, 50 μm (B) F4/80 mRNA levels in livers; n = 5 per group (C) ELISA analysis of serum TNFα levels; n = 5–6 per group (D) Relative mRNA expression of pro-inflammation genes in mouse liver; n = 3–5 per group. β-ACTIN was used as an internal control for normalizing the mRNA levels. Data are statistically analysed as means ± SEM. # p < 0.05, MCD compared with the MCS group of mice. *p < 0.05, MCD compared with the MCD + VTE group. NS, no significance.

FIGURE 2 | Effect of WMW on DSS-induced acute colitis in mice.(E) Double immunofluorescent signals from F4/80 and TNF-α, F4/80 Immunohistochemistry in DSS-induced colitis tissue at different groups (original magnification, 400 ×). n = 7–10, means ± SEM.*p <0.05, **p < 0.01,***p < 0.001.