Nrf2 Antibody - #AF7006

Figure 2. Nrf2 significantly suppresses TLR4 and IRF1 levels in a mouse model of CaOx nephrocalcinosis. (A) IHC staining of kidney Nrf2, TLR4, and IRF1 in CaOx nephrocalcinosis mice treated with SFN (400× magnification; scale bar: 40 µm). SFN treatment increases Nrf2 and inhibits the expression of TLR4 and IRF1 in CaOx nephrocalcinosis mouse tubular epithelial cells. (B) Quantification of IHC staining of Nrf2, TLR4, and IRF1 in SFN treated CaOx nephrocalcinosis mouse model. (C) qPCR detection of Nrf2, TLR4, and IRF1 expression in SFN-treated CaOx nephrocalcinosis mouse kidney samples and comparison with normal controls. (D, E) Pearson’s correlation analysis of Nrf2 levels relative to TLR4 and IRF1. Data represent the mean ± standard error (SE) of three independent experiments. *P < 0.05; **P < 0.01, as determined by one-way ANOVA (B, C) or Pearson’s correlation (D, E).

Figure 3. Nrf2 suppresses TLR4 and IRF1 levels and promotes M2Mϕ polarization in vitro. (A) BMDM and COM-TECs coculture schematic diagram. Western blot (B) and qPCR (C) analyses of Nrf2, TLR4, IRF1, iNOS, and ARG-1 levels in BMDMs co-cultured with increasing COM dose stimulated TECs. GAPDH was used as an internal control. (D, F) Western blot detection of Nrf2, TLR4, IRF1, iNOS, and ARG-1 levels following SFN treatment or Nrf2 upregulation/downregulation in BMDMs co-cultured with COM-stimulated TECs. GAPDH was used as an internal control. (E, G) The distribution of iNOS (green) and ARG-1 (red) in BMDMs according to immunofluorescence. (H, I) Flow cytometric analysis of the polarization state of BMDMs using anti-CD11c and anti-CD206 in F4/80+ and CD11b+ cells. The data are shown as the mean ± standard error (SE) of three independent experiments. *P < 0.05; **P < 0.01, as determined by Student’s t test (C) or one-way ANOVA (E, G, H, I).

Fig. 5. Effect of treatments of MGO, Que-mono-MGO, and Que-di-MGO on the expression levels of apoptotic markers and components of AKT and Nrf2-HO-1/NQO-1 signaling pathways. Significant differences (p < 0.05) between samples of different treatments are marked with different letters on each column.

Fig. 2. Pretreatment with monotropein protected against renal oxidative stress induced by cisplatin. The level of serum GSH(A). The level of serum MDA(B). The level of serum SOD(C). The level of serum CAT(D). The protein expressions of Nrf2, HO-1 and NQO1 (E) were detected by Western blot in kidney tissue. The protein expressions of Nrf2 (F), HO-1 (G) and NQO1 (H) were quantitated by Image software. Data are showed as mean ± S.E.M of 6 mice in each group. #P < 0.05, ##P < 0.01 vs. Normal group; *P < 0.05, **P < 0.01 vs. cisplatin group.

FIGURE 6 | DCL enhances the activation of Keap1/Nrf2/HO-1 in LPS/IFNγ-stimulated RAW264.7 macrophages. RAW264.7 cells were treated with DCL (1, 3, and 9 μM) or curcumin (5 μM) for 1 h, and followed by LPS/IFNγ incubation for additional 8 h. The protein expression of Keap1 (A), Nrf2 (C), and HO-1 (G) were measured by western blot.

FIGURE 5 | Expression of the targeted proteins in mice liver. Values are shown as mean ± SD (n = 8). Data (mean ± SD) with different case letters are statistically different with each other at LSD multiple comparisons, which with red letters differed significantly as compared to model group level. uppercase letter p < 0.01 vs.another group.

FIGURE 4 | Immunohistochemistry observation of nuclear factor erythroid 2-related factor 2 (Nrf2) expressions in livers of mice. Images were obtained at 400×magnification (scale bar=50mm).