Journal: Cancers

Article Title: Study of Biological Effects Induced in Solid Tumors by Shortened-Duration Thermal Ablation Using High-Intensity Focused Ultrasound

doi: 10.3390/cancers16162846

Figure Lengend Snippet: Immunolabeling for IL-1α (red, ( left ) panel) and IL-1β (green, ( right ) panel) in a HIFU-treated tumor (3 days post-sonication) as compared to an untreated control tumor. White arrows indicate immunopositive cells. Bisbenzimide was used for visualization of cell nuclei (blue). Histograms show the intensity of immunolabeling for IL-α and IL1-β per microscopic field of view. The intensity of immunoreactivity is presented as the height of the surface plot profiles.

Article Snippet: Then, the sections were incubated for 1 h at 37 °C with PBS+T containing 1% normal goat serum and one or two of the following primary antibodies (Abs): (1) murine monoclonal Ab against vascular endothelial growth factor (VEGF) (Santa Cruz Biotechnology, Santa Cruz, CA, USA, dil. 1:400); (2) rabbit polyclonal Ab against IL 1-β (Invitrogen-Molecular Probes, Eugene, OR, USA, dil. 1:400); (3) murine monoclonal Ab against IL 1-α (Santa Cruz Biotechnology, CA, USA, dil. 1:400); (4) murine monoclonal Ab against TNF-α (Santa Cruz Biotechnology, CA, USA, dil. 1:400); (5) murine monoclonal Ab against macrophages (Santa Cruz Biotechnology, CA, USA, dil. 1:400); (6) murine monoclonal Ab against lymphocytes CD8 (DAKO, Santa Clara, CA, USA, dil. 1:400), and (7) murine monoclonal Ab against INF-γ (Santa Cruz Biotechnology, CA, USA, dil. 1:400).

Techniques: Immunolabeling, Sonication, Control

Journal: Mediators of Inflammation

Article Title: Chrysophanol Inhibits NALP3 Inflammasome Activation and Ameliorates Cerebral Ischemia/Reperfusion in Mice

doi: 10.1155/2014/370530

Figure Lengend Snippet: Model of danger signals activation of the NALP3 inflammasome. Tissue injury leads to the formation and release of danger signals such as ATP or uric acid crystals that are recognized by the innate immune system. A number of these signals mediate a potassium efflux or other secondary intracellular danger signals that are required for NALP3 inflammasome activation [ , ]. NALP3 inflammasome then oligomerizes to recruit the adaptor ASC and caspase-1 . Activation of caspase-1 results in the processing and maturation of pro IL-1 β into its biologically active form, active IL-1 β [ , ]. Active IL-1 β will then trigger the IL-1 β receptor, leading to the activation of multiple cytokines involved in the inflammation cascade .

Article Snippet: Brain sections (5 μ m thick) were blocked in 3% H 2 O 2 , 3% normal goat serum, and then incubated overnight with anti-mouse NALP3, ASC and IL-1 β rabbit polyclonal antibodies (1 : 100, Abcam Biotechnology), and anti-mouse caspase-1 rabbit polyclonal antibody (1 : 100, Sigma Biotechnology).

Techniques: Activation Assay

Journal: Mediators of Inflammation

Article Title: Chrysophanol Inhibits NALP3 Inflammasome Activation and Ameliorates Cerebral Ischemia/Reperfusion in Mice

doi: 10.1155/2014/370530

Figure Lengend Snippet: Cellular distribution of NALP3 inflammasome components and IL-1 β in the ischemic penumbra zone of cerebral cortex. Sections were triple-stained for NALP3, ASC, caspase-1 and IL-1 β (red), NeuN (green), and nucleus (blue). In tMCAO group, the neuronal karyopyknosis and chromatin margination could be observed and there was a small amount of neutrophils in marginal zone of infarction. In the normal brain (Sham), caspase-1 and NALP3 were mainly present in the nucleus of neurons whereas IL-1 β was mainly localized in the cytoplasm. ASC was present in both the nucleus and cytoplasm. After tMCAO, NALP3 and caspase-1 were redistributed predominantly to the cytoplasm, while ASC and IL-1 β location remained unchanged.

Article Snippet: Brain sections (5 μ m thick) were blocked in 3% H 2 O 2 , 3% normal goat serum, and then incubated overnight with anti-mouse NALP3, ASC and IL-1 β rabbit polyclonal antibodies (1 : 100, Abcam Biotechnology), and anti-mouse caspase-1 rabbit polyclonal antibody (1 : 100, Sigma Biotechnology).

Techniques: Staining

Journal: Mediators of Inflammation

Article Title: Chrysophanol Inhibits NALP3 Inflammasome Activation and Ameliorates Cerebral Ischemia/Reperfusion in Mice

doi: 10.1155/2014/370530

Figure Lengend Snippet: The dynamic expression of NALP3 inflammasome and IL-1 β during the natural inflammatory response induced by tMCAO. Western blot of dynamic expression ((a), (b), (c)). Bar graph illustrating the dynamic expression of NALP3, ASC, active caspase-1, and active IL-1 β in brain tissues (d). Bar graph illustrating dynamic expression of NALP3, ASC, caspase-1, and IL-1 β as determined by immunohistochemistry (e). Bar graph illustrating the mRNA dynamic expression of NALP3, ASC, caspase-1, and IL-1 β (f). For NALP3, ☆ P < 0.05 versus Sham, ★ P < 0.05 versus Sham, 3 h, 6 h, 12 h, 48 h, and 72 h. For ASC, △ P < 0.05 versus Sham. For active caspase-1 and active IL-1 β , ◊ P < 0.05 and □ P < 0.05 versus Sham, ◆ P < 0.05 and ■ P < 0.05 versus Sham, 3 h, 6 h, 12 h, and 72 h.

Article Snippet: Brain sections (5 μ m thick) were blocked in 3% H 2 O 2 , 3% normal goat serum, and then incubated overnight with anti-mouse NALP3, ASC and IL-1 β rabbit polyclonal antibodies (1 : 100, Abcam Biotechnology), and anti-mouse caspase-1 rabbit polyclonal antibody (1 : 100, Sigma Biotechnology).

Techniques: Expressing, Western Blot, Immunohistochemistry

Journal: Mediators of Inflammation

Article Title: Chrysophanol Inhibits NALP3 Inflammasome Activation and Ameliorates Cerebral Ischemia/Reperfusion in Mice

doi: 10.1155/2014/370530

Figure Lengend Snippet: Immunohistochemical staining of NALP3, ASC, caspase-1, and IL-1 β in the cerebral cortex at 24 h after ischemia (400x magnification). As compared with tMACO and Vehicle groups, the expression of NALP3, caspase-1, and IL-1 β was significantly reduced in CHR-H and CHR-M groups, but the expression of ASC remained unchanged.

Article Snippet: Brain sections (5 μ m thick) were blocked in 3% H 2 O 2 , 3% normal goat serum, and then incubated overnight with anti-mouse NALP3, ASC and IL-1 β rabbit polyclonal antibodies (1 : 100, Abcam Biotechnology), and anti-mouse caspase-1 rabbit polyclonal antibody (1 : 100, Sigma Biotechnology).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Mediators of Inflammation

Article Title: Chrysophanol Inhibits NALP3 Inflammasome Activation and Ameliorates Cerebral Ischemia/Reperfusion in Mice

doi: 10.1155/2014/370530

Figure Lengend Snippet: Effect of CHR on protein expression of NALP3, ASC (a), caspase-1, active caspase-1 (b), pro IL-1 β and active IL-1 β (c). For NALP3, active caspase-1 and active IL-1 β , ★ P < 0.05, ◆ P < 0.05 and ■ P < 0.05 versus tMCAO and Vehicle (d). Bar graph of immunohistochemistry illustrating the expression of NALP3 ( ☆ P < 0.05 versus tMCAO, Vehicle and CHR-M, ★ P < 0.05 versus tMCAO, Vehicle), ASC, caspase-1 ( ◊ P < 0.05 versus tMCAO, Vehicle and CHR-M, ◆ P < 0.05 versus tMCAO, Vehicle) and IL-1 β ( ■ P < 0.05 versus tMCAO and Vehicle) (e). Bar graph illustrating CHR reduced to mRNA of NALP3, caspase-1 and IL-1 β ( ★ P < 0.05, ◆ P < 0.05 and ■ P < 0.05 versus tMCAO and Vehicle) (f).

Article Snippet: Brain sections (5 μ m thick) were blocked in 3% H 2 O 2 , 3% normal goat serum, and then incubated overnight with anti-mouse NALP3, ASC and IL-1 β rabbit polyclonal antibodies (1 : 100, Abcam Biotechnology), and anti-mouse caspase-1 rabbit polyclonal antibody (1 : 100, Sigma Biotechnology).

Techniques: Expressing, Immunohistochemistry