Fig 2: TLR4 deficiency reduced macrophage accumulation in diabetic nephropathy.Macrophage (CD68+ or F8/40+) accumulation was evident in WT mice with diabetes, but prevented by TLR4 deficiency in both the interstitial (A) and glomerular (B & C) compartments. Data are present as means ± SEM; * p<0.05; *** p<0.001.

Fig 3: Expression of TLR4 and its endogenous ligands was upregulated in the early diabetic kidney in WT mice.mRNA expression of TLR4 and its ligands in kidney was upregulated at 10 weeks after diabetes induction (A & B). The data are presented as means ± SD; * p<0.05; ** p<0.01; *** p<0.001. n = 7 per group. (C) Representative sections of kidney were stained with a rabbit anti-mouse-TLR4 polyclonal antibody in the top panels and a goat anti-mouse-TLR4 polyclonal antibody in the bottom panels.

Fig 4: Endometriosis upregulated TLR4 in nociceptors. a–h Double immunostaining showed co-expression of TLR4 with nociceptive marker, CGRP, IB4, and TRPV1 in rat DRG from sham and EM 14d groups. Scale bar 50 µm. i Parentages of TLR4+ cells in neurons with nociceptive markers (IB4, CGRP, and TRPV1) were increased in the EM 14d group compared with that in sham group. n > 100 neurons from 3 rats in each group, *p < 0.05, EM 14d vs sham, chi-square test

Fig 5: Effect of AG on IL-1ß-induced activation of the TLR4/MyD88/NF-?B signaling pathway in human NP cells. Cells were stimulated by IL-1ß with or without AG treatment. (A) The levels of TLR-4, MyD88, p65 and p-p65 were determined by western blot analysis. (B) Quantification of TLF4 levels. (C) Quantification of MyD88 levels. (D) Quantification of p65/p-p65 ratio. (E) p65 expression was also measured by immunofluorescence (magnification, ×400). The values are presented as the mean ± standard deviation of 3 independent experiments. *P<0.05 vs. control group; #P<0.05 vs. IL-1ß group. AG, andrographolide; IL-1ß, interleukin 1ß; TLR-4, toll-like receptor 4; MyD88, myeloid differentiation primary response protein MyD88; p65, Transcription factor p65; p, phosphorylated.