Fig 1: The effect of inhibiting signalling of the IL-1 pathway on growth of E0771 cells in mammary glands and spontaneous metastasis to lung and bone (A) Protocol outline for orthotopic injection of E0771 cells into mammary glands and administration of PBS (control), MLX01, VX765 or Anakinra in mice. (B) Primary tumour volume following daily administration of PBS (control), MLX01, VX765 or Anakinra. (C) Survival curve (D) IVIS images of dissected femur/tibia ex vivo following exposure to D-luciferin for 5 min. (E) Percentage of mice that developed spontaneous bone metastasis and size of tumours in femurs/tibiae as assessed ex vivo by IVIS. (F) IVIS images of dissected lungs, ex vivo (G) Percentage of mice that developed spontaneous lung metastasis and size of tumours in lungs as assessed by IVIS, ex vivo. The graphs represent mean ± SD, * p < 0.05, ** p < 0.01, **** p < 0.0001.
Fig 2: Inhibiting IL-1 signalling with IL1ß inhibitor MLX01, Caspase-1 inhibitor VX765 and IL1R antagonist Anakinra reduced metastatic outgrowth from E0771 cells disseminated in the bone. (A) Outline of experimental protocol for administration of PBS (control), MLX01, VX765 and Anakinra in mice following intra-cardiac injection of E0771. (B) Representative whole body IVIS images of mice at Day 8 and Day 15 post injection of E0771 cells. (C) Numbers of overt metastases per mouse 8-, 15- and 16-days post injection of E0771 cells following daily administration of PBS (control), MLX01, VX765 or Anakinra. (D) Total luciferase expression (photons per second (P/S)) of whole mice 8 and 15 days after E0771 cell injection. (E) Numbers of mice that developed bone metastases 8, 15 and 16 days after E0771 cells had been disseminated in the bone following daily administration of PBS (control) MLX01, VX765 or Anakinra. (F) Size of tumours that grew in the bone as assessed by photons per second (P/S) in luciferase expressing E0771 tumour cells. (G) Shows effects of PBS (control), MLX01, VX765 or Anakinra on metastatic outgrowth of tumours following dissemination into the lungs, kidneys and livers. (H) Protocol outline for administration of Pacritinib in mice after intra-cardiac injection of E0771. (I) Representative images of whole body IVIS imaging 17 days post E0771 injection. (J) Numbers of overt metastases per mouse 17 days post injection of E0771 cells and following daily administration of PBS (control), or Pacritinib. (K) Total luciferase signal from E0771 tumour cells (photons per second (P/S)) in whole mice 17 days following injection of E0771 cells and administration of PBS (control) or Pacritinib. (L) Numbers of mice that developed bone metastases 17 days after E0771 cells had been disseminated in the bone following daily administration of PBS (control) or Pacritinib and size of tumours that grew in the bone as assessed by IVIS imaging in (M). (N) Shows effects of PBS (control), or Pacritinib on metastatic outgrowth of tumours following dissemination into the lungs, kidneys and livers. *** p < 0.001, **** p < 0.0001.
Fig 3: The schematic hypothesis of the function of LLLT to promote wound healing. Our results suggest that LLLT decreases IL-ß and IL-6, and increases IL-1RA at the gingival wound, promoting wound healing and bone regeneration. The IL-1 signaling pathway is initiated when IL-1a or IL-1ß binds to the members of the IL-1R1 family, which recruits MyD88, IRAK, and TRAF6 to activate NF-?B and MAPK and then controls the transcription of several inflammatory genes. IL-1RA can suppress this interaction competitively
Fig 4: NTB adjuvant promote the immature DCs maturation and cytokines Secretion. (A) Antigen uptake of PBS, TBI, BNE and NTB formulated with GFP antigen by immature DCs. The DCs were imaged by CLSM after being incubated with naïve GFP protein for 30min. RED (Phalloidin-stained cell cytoskeleton), Green (GFP fluorescence) and Blue (DAPI-stained cell nucleus). Scale bar=20µm. (B–D) Effects of PBS, TBI, BNE and NTB-formulated antigen on the activation of immature DCs. The DCs were stimulated with PBS, TBI, BNE and NTB-formulated antigen for 24h, and supernatants were harvested for cytokine detection. Concentrations of cytokine (IL-1b, IL-6, and TNF-a) were detected by ELISA. The results are shown as the means ± SD. *P <0.05; **P < 0.01; ***P < 0.001.
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