Fig 1: Elevated ICP increases IL-18 expression in microglia following IR in vivo. (A) Immunoreactive bands of IL-18 (22 kDa) and ß-actin (43 kDa). (B) Protein expression levels of IL-18 were significantly increased in the IR group and the IR + NS group compared with the sham group. There was no significant difference between the IR group and the IR + NS group. The levels in the IR + HS group were significantly lower than the IR group when ICP levels were reduced by HS. (C) Immunofluorescence images showing the expression of (C-a, C-e, C-i and C-m) Iba1+ microglia (green), (C-b, C-f, C-j and C-n) IL-18 (red), (C-c, C-g, C-k and C-o) the co-localization of IL-18 and microglia, and (C-d, C-h, C-l and C-p) high amplification images of the microglial cells in peri-ischemic cortex. Increased IL-18 immunofluorescence was observed in the IR group and the IR + NS group compared with the sham group. In the IR + HS group, IL-18 fluorescence was notably reduced. Scale bar, 20 µm. n=4 per group. **P<0.01 vs. sham group; ##P<0.01 vs. IR group. ICP, intracranial pressure; sham, sham-operated; IR, ischemia-reperfusion; NS, normal saline; HS, hypertonic saline group; ns, non-significant.
Fig 2: Upregulation of TRPV4 in the SN aggravated inflammation in PD mice. After infused with 0.3 µl AAV-TRPV4 bilaterally into the SN of mice for 21 days and administrated with MPTP at 30 mg/kg/day for 7 days, the SN was rapidly dissected out. The protein levels of procaspase-1, IL-18, COX-2 and 5-LOX in the SN of mice were detected by Western blot. A Upregulation of TRPV4 further decreased the MPTP-induced the reduction in procaspase-1 in the SN (n = 6). B Upregulation of TRPV4 further increased the MPTP-induced high level of IL-18 in the SN (n = 6). C Upregulation of TRPV4 further increased the MPTP-induced high level of COX-2 in the SN (n = 6). D Upregulation of TRPV4 further increased the MPTP-induced high level of 5-LOX in the SN (n = 6). All data are presented as mean ± SEM. Statistical significance: *P < 0.05, **P < 0.01, ***P < 0.001
Fig 3: Effect of RKC-B1 on NLRP3/cleaved caspase-1 signaling pathway. (a) Protein expression levels of NLRP3; (b) protein expression levels of cleaved caspase-1; (c) protein expression levels of IL-18. Data are expressed as the mean ± SD (n = 5/group). Statistical significance is assessed by one-way ANOVA analysis to compare the results between different groups * p < 0.05, ** p < 0.01 vs. LPS model group.
Fig 4: Treatment with IL-18BP decreased free IL-18 levels in mouse serum. Mice received single dose of rhIL-18BP (2 mg/kg) or vehicle-control SC injection 48 h post-9.4 Gy TBI and serum were collected on day 3 and 4 after TBI. Levels of mouse IL-18, endogenous mouse IL-18BP and exogenous human IL-18BP in mouse serum were measured with corresponding ELISA kits. (a) The total levels of IL-18BP (endogenous mouse IL-18BP and exogenous human IL-18BP) in vehicle-treated or rhIL-18BP-treated mouse serum and (b) free IL-18 in mouse serum. Data of total IL-18 in serum and free IL-18 levels by calculation in vehicle treated and IL-18BP treated group 3 and 4 day after TBI are included. Results were obtained from two independent experiments (6 mice/group/experiment, N = 12). Means ± SD. *p < 0.05; **p < 0.01; IL-18BP-treated vs. vehicle-treated.
Fig 5: Immunohistochemical (IHC) staining of bone marrow with anti-IL-18 and IFN-? antibodies. Mice were irradiated with 9.4 Gy TBI and then treated with vehicle or IL-18BP at 48 h post TBI. Mice were sacrificed at day (d) 3, d4 and d7 after TBI. Un-irradiated mice were included for comparison. Sternum bone marrow specimens were stained with IL-18 (a), IFN-? (b) or rabbit IgG (c) antibodies. Brown color indicates positive staining. Scale bar = 100 µm. Quantification of IL-18 and IFN-? IHC staining was shown in (d), N = 3.*p < 0.05. Note that the d3 vehicle treated group was significantly different with any other group in the IFN-? staining.
Supplier Page from Abcam for Anti-IL-18 antibody [EPR19956]