Phospho-IRAK1 (Thr387) Antibody - #AF8009

Fig. 5 S100A14 promotes ubiquitin–proteasome-mediated degradation of IRAK1. a Immunoblotting analysis of P-IRAK1 (T387) and IRAK1 protein levels in the S100A14 overexpressing cells and S100A14 knocked-down cells.

Figure 7. FCN-A promoted the M1 polarization of BMDMs through a TLR4/MyD88-dependent pathway in vitro. (a) The protein expressions of the purified GST-FCN-A and GST were determined by SDS-PAGE. (b) The extracted membrane proteins from RAW264.7 cells were incubated with the purified GST-FCN-A or GST proteins. Co-IP analysis of the interaction between TLR4 of macrophage and GST-FCN-A was performed by using anti-TLR4. Rabbit IgG was used as a negative control in co-IP. (c, e) BMDMs, isolated from WT, TLR4-/- or MyD88-/- mice, were stimulated with FCN-A (10g/mL) for 24 h, then the expressions of iNOS and Arg-1 from BMDMs were examined by Western blot analysis. (d, f) The levels of pro-inflammatory cytokines IL-1in cell lysates, and secreted IL-6, TNF- were detected by ELISA. (g, h) Western blot analysis of p-IRAK1, p-p65, p-ERK1/2, and p-JNK in the BMDM lysates of TLR4-/-, MyD88-/- or WT after stimulation with FCN-A for 0-45 min. In d and f, values are mean ± [SEM] from three independent experiments.

Fig. 4. LPS-induced activation of the IRAK-4/IRAK-1/NF-κB signalling pathway in OPCs was inhibited by ART. a, b, c, d Western blotting was used to quantify the protein levels of p-IRAK-4, IRAK-4, p-IRAK-4/IRAK-4, p-IRAK-1, IRAK-1, p-IRAK-1/IRAK-1, phospho-NF-kB p65, NF-κB p65, and phospho-NF-kB p65/NF-κB p65 in 4 groups (the Ctrl, ART, LPS, and LPS + ART groups). The data are expressed as the means ± SEMs (n = 6/group) (*p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001).

Fig. 5. ART reduces NF-κB nuclear aggregation after LPS stimulation. a, b, c Immunofluorescence was used to compare the relative fluorescence of p-IRAK-4 (red) and p-IRAK-1 (red) in OPCs across several groups (the Ctrl, ART, LPS, and LPS + ART groups), and the proportion (%) of nuclear NF-κB p65 (red) was determined. DAPI staining is shown in blue. Scale bar = 50 μm. The data are presented as the means ± SEMs (n = 6). ****P < 0.0001 versus the indicated groups.

Fig. 8. Effects of ONSMP on JNK1/2 signaling pathway. (A) Representative western blotting bands of p-IRAK4, IRAK4, p-IRAK1, IRAK1, p-TAK1, TAK1, p-MKK4, MKK4, p-MKK7, MKK7, p-JNK1/2, JNK1/2, MD2, TLR4, MyD88, and c-Jun in the myocardium, with vinculin as internal reference. (B–K) Statistical graphs of p-IRAK4/IRAK4, p-IRAK1/IRAK1, p-TAK1/TAK1, p-MKK4/MKK4, p-MKK7/MKKK7, p-JNK1/2/JNK1/2, MD2, TLR4, MyD88, and c-Jun (n = 3). Data are presented as mean ± SEM. nsP > 0.05; ▲▲P < 0.01 vs the sham; *P < 0.05, **P < 0.01 vs the model; #P < 0.05, ##P < 0.01 vs the ONSMP-L; ※P < 0.05, ※※P < 0.01 vs the ONSMP-M.

Fig. 4. Effects of INR on Tollip expression and mitochondrial damage in kidney tissues of PQ-intoxicated rats. (A) Tollip expression in kidney tissues was detected by immunohistochemistry. Scale bars: 200 μm (100× magnification) and 50 μm (400× magnification). (B, C) Western blot showed the expression of Tollip, p-IRAK1 and IRAK1 in kidney tissues. (D, E) Western blot evaluated the release of Cytochrome C from mitochondria into cytosol. ###p < 0.001 vs. PQ.