Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Alleviation of chronic pain following rat spinal cord compression injury with multimodal actions of huperzine A

doi: 10.1073/pnas.1300083110

Figure Lengend Snippet: Antibodies used in immunocytochemical assays

Article Snippet: NK1R , Neurokinin-1 receptor , Goat , 1:250 , Santa Cruz Biotechnology , Santa Cruz, CA.

Techniques: Activation Assay, Expressing, Dominant Negative Mutation, Marker, Activity Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Enhancement of innate and adaptive anti-tumor immunity by serum obtained from vascular photodynamic therapy-cured BALB/c mouse

doi: 10.1007/s00262-021-02917-4

Figure Lengend Snippet: Actions of complement C5a on cytokine release. (a) Cytokine induction. The serum from VPDT-cured mice was first treated either by heat (Heat) or heparin (Heparin) (20 U/ml) before incubation with the CT26 cells for 24 h. The concentrations of the cytokines were measured (means ± S.E.M., N = 3). (b) Concentration of C5a in the serum of different groups of mice. The “Anti-C5a” group refers to the serum from VPDT-cured mice in which anti-C5a (2 μg/ml) was added. C5a was determined using ELISA (means ± S.E.M., N = 3). (c) Cytokine expression (± anti-C5a/ recombinant C5a). CT26 cells were incubated with either the serum from VPDT-cured mice (Cured) with and without anti-C5a or heat-treated serum from VPDT-cured mice (Heat) with and without recombinant C5a (2 μg/ml) for 24 h before the level of IL-6 was determined (means ± S.E.M., N = 3). (d) CD88 expression. After incubating CT26 cells with various sera, the level of surface CD88 expression was determined using flow cytometry (means ± S.D., N = 3). (e) Cytokine expression (± antagonist). CT26 cells were incubated with the serum from VPDT-cured mice in the presence of different concentrations of W-54011 for 24 h before the concentration of IL-6 in the supernatant was measured (means ± S.E.M., N = 3). A concentration of 20 μM for W-54011 was used in the study on the mRNA levels of CXCL1–3 (means ± S.E.M., N = 3). The data obtained from ELISA and real-time PCR were analyzed using either Welch’s t test or one-way ANOVA and those from flow cytometry were analyzed using one-way ANOVA test. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; n.s., not significant

Article Snippet: The C5a receptor antagonist W-54011(Santa Cruz Biotechnology, sc-203863) and recombinant C5a and C3a (Sigma, SRP4895A, R&D Systems, 8085–C3–025) were also examined for their effects on cytokine release.

Techniques: Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing, Recombinant, Flow Cytometry, Real-time Polymerase Chain Reaction

Journal: Frontiers in Pharmacology

Article Title: The NLRP3 inflammasome is involved in resident intruder paradigm-induced aggressive behaviors in mice

doi: 10.3389/fphar.2023.974905

Figure Lengend Snippet: Behavioral phenotype of mice in each group. (A) Schematic representation of the experimental design. (B) Open field test representative trajectory graph. (C) Elevated plus-maze test representative trajectory graph. (D) Open field test results. (E) Elevated plus-maze test results. (F) Attack behavior test results. Data are expressed as means ± SEMs. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Control, producers of non-aggressive behavior after stress; Model, aggressive behavior after stress; A804598, P2X7R antagonist; IL-1Ra, IL-1β blocker; NLRP3 −/− , NLRP3 knockout mice. n = 8/group. (TIFF format, 1200 dpi, 2-column fitting image).

Article Snippet: Chemicals are listed as follows: A804598 (HY-100483, MedChemExpress Co., Ltd. United States); IL-1Ra (HY-P7029A, MCE, United States); DMSO (D8370, Solarbio, China); Mouse NLRP3 ELISA kit (JYM0765Mo, Wuhan ColorfulGene biological technology Co., Ltd. China); Mouse IL-1β ELISA kit (JYM0531Mo, Wuhan ColorfulGene biological technology Co., Ltd. China); BCA Protein Assay Kit (PC0020, Solarbio, China); SDS-PAGE Gel Rapid Preparation Kit (G2037-50T, Servicebio, China); Recombinant Anti-beta Actin antibody (GB15003, Servicebio, China); NLRP3 Antibody (DF7438, Affinity Biologicals, Canada); Recombinant Anti-P2X7 antibody (ab259942, abcam, United Kingdom); HRP-conjugated goat anti-rabbit IgG (GB23204, Servicebio, China); Anti-Iba1 Rabbit pAb (GB11105, Servicebio, China); Cy3-labeled goat anti-rabbit IgG (GB21303, Servicebio, China); FITC-labeled goat anti-rabbit IgG (GB22303, Servicebio, China); Anti-BrdU Mouse mAb (GB12051, Servicebio, China); Anti-NeuN Rabbit pAb (GB11138, Servicebio, China); HE staining solution (G1005-100ML, Servicebio, China).

Techniques: Control, Knock-Out

Journal: Frontiers in Pharmacology

Article Title: The NLRP3 inflammasome is involved in resident intruder paradigm-induced aggressive behaviors in mice

doi: 10.3389/fphar.2023.974905

Figure Lengend Snippet: The P2X7R was activated in the resident intruder paradigm. (A) Representative images of P2X7R (green) and DAPI (blue) labeling in the hippocampus CA1, CA3, and DG after drug interventions; scale bar = 200 and 50 µm. (B) Positive staining area of P2X7R and DAPI double-labeled (P2X7R + DAPI) proteins in the hippocampus CA1 (** p < 0.01 Control vs. Model ; n = 3/group), CA3 (** p < 0.01 Control vs. Model ; * p < 0.05 A804598 vs. Model ; n = 3/group), and DG (** p < 0.01 Control vs. Model ; * p < 0.05 A804598 vs. Model ; n = 3/group). (C) Representative immunoreactive bands showing the protein levels of hippocampal P2X7R in the Control, Model, A804598, IL-1Ra, and NLRP3 −/− mice. (D) Statistical results show that A804598, IL-1Ra, and NLRP3 −/− decreased the protein expression of P2X7R ( n = 3, **** p < 0.0001 vs. Model).

Article Snippet: Chemicals are listed as follows: A804598 (HY-100483, MedChemExpress Co., Ltd. United States); IL-1Ra (HY-P7029A, MCE, United States); DMSO (D8370, Solarbio, China); Mouse NLRP3 ELISA kit (JYM0765Mo, Wuhan ColorfulGene biological technology Co., Ltd. China); Mouse IL-1β ELISA kit (JYM0531Mo, Wuhan ColorfulGene biological technology Co., Ltd. China); BCA Protein Assay Kit (PC0020, Solarbio, China); SDS-PAGE Gel Rapid Preparation Kit (G2037-50T, Servicebio, China); Recombinant Anti-beta Actin antibody (GB15003, Servicebio, China); NLRP3 Antibody (DF7438, Affinity Biologicals, Canada); Recombinant Anti-P2X7 antibody (ab259942, abcam, United Kingdom); HRP-conjugated goat anti-rabbit IgG (GB23204, Servicebio, China); Anti-Iba1 Rabbit pAb (GB11105, Servicebio, China); Cy3-labeled goat anti-rabbit IgG (GB21303, Servicebio, China); FITC-labeled goat anti-rabbit IgG (GB22303, Servicebio, China); Anti-BrdU Mouse mAb (GB12051, Servicebio, China); Anti-NeuN Rabbit pAb (GB11138, Servicebio, China); HE staining solution (G1005-100ML, Servicebio, China).

Techniques: Labeling, Staining, Control, Expressing

Journal: Frontiers in Pharmacology

Article Title: The NLRP3 inflammasome is involved in resident intruder paradigm-induced aggressive behaviors in mice

doi: 10.3389/fphar.2023.974905

Figure Lengend Snippet: Correlations between the expression levels of NLRP3 in the hippocampus or serum and aggressive behaviors. (A) Representative immunoreactive bands showing the protein levels of hippocampal NLRP3 in the Control, Model, A804598, IL-1Ra, and NLRP3 −/− mice. (B) Statistical results show that A804598, IL-1Ra, and NLRP3 −/− decreased the protein expression of NLRP3 ( n = 3, **** p < 0.0001 vs. Model). (C) NLRP3 and IL-1βhippocampus and serum levels. (D) Pearson correlation analyses were performed by comparing the expression levels of NLRP3 in the hippocampus or serum and aggressive behavior latency or aggressive behavior score. Correlations were performed considering some animals ( N = 3). The coefficient r and p -value for each correlation are presented in a box. Statistically significant correlations are highlighted in red ( p < 0.05). Data are expressed as means ± SEMs. Control, producers of non-aggressive behavior after stress; Model, aggressive behavior after stress; A804598, P2X7R antagonist; IL-1Ra, IL-1β blocker; NLRP3 −/− , NLRP3 knockout mice. (TIFF format, 1200 dpi, 2-column fitting image).

Article Snippet: Chemicals are listed as follows: A804598 (HY-100483, MedChemExpress Co., Ltd. United States); IL-1Ra (HY-P7029A, MCE, United States); DMSO (D8370, Solarbio, China); Mouse NLRP3 ELISA kit (JYM0765Mo, Wuhan ColorfulGene biological technology Co., Ltd. China); Mouse IL-1β ELISA kit (JYM0531Mo, Wuhan ColorfulGene biological technology Co., Ltd. China); BCA Protein Assay Kit (PC0020, Solarbio, China); SDS-PAGE Gel Rapid Preparation Kit (G2037-50T, Servicebio, China); Recombinant Anti-beta Actin antibody (GB15003, Servicebio, China); NLRP3 Antibody (DF7438, Affinity Biologicals, Canada); Recombinant Anti-P2X7 antibody (ab259942, abcam, United Kingdom); HRP-conjugated goat anti-rabbit IgG (GB23204, Servicebio, China); Anti-Iba1 Rabbit pAb (GB11105, Servicebio, China); Cy3-labeled goat anti-rabbit IgG (GB21303, Servicebio, China); FITC-labeled goat anti-rabbit IgG (GB22303, Servicebio, China); Anti-BrdU Mouse mAb (GB12051, Servicebio, China); Anti-NeuN Rabbit pAb (GB11138, Servicebio, China); HE staining solution (G1005-100ML, Servicebio, China).

Techniques: Expressing, Control, Knock-Out