Journal: American Journal of Translational Research

Article Title: Silencing of TLR7 protects against lipopolysaccharide-induced chondrocyte apoptosis and injury by blocking the p21-mediated JAK2/STAT3 pathway

doi:

Figure Lengend Snippet: Production of inflammatory mediators and cytokines in LPS-stimulated chondrocytes were reduced by TLR7 knockdown. Chondrocytes were transfected with si-NC and si-TLR7 for 48 h and then cells were treated with 5 μg/ml LPS for 12 h. A. The content of NO was detected in cells. B. The protein level of iNOS in cells was analyzed by western blot assay. C. mRNA expressions of IL-1β, TNF-α, and IL-6 in chondrocytes were assessed by RT-qPCR. D-F. The contents of IL-1β, TNF-α, and IL-6 in chondrocytes were detected through ELISA assay. Columns present the mean ± SEM (n≥3); *P<0.05 versus control; #P<0.05 versus LPS group.

Article Snippet: ELISA assay The corresponding ELISA kits were used to measure the contents of IL-6, TNF-α and IL-1β (BioSite, Paris, France).

Techniques: Transfection, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Programmable Stapling Peptide Based on Sulfonium as Universal Vaccine Adjuvants for Multiple Types of Vaccines

doi: 10.1002/advs.202409567

Figure Lengend Snippet: The efficacy evaluation of M‐CP as a universal adjuvant for influenza vaccine and herpes zoster vaccine. a) Schematic illustration of the vaccination plan to evaluate the universality of M‐CP for different vaccines. Balb/c mice were intramuscularly injected with different formulations as planned. b,c) The anti‐BY antibody titer at b) day 28 and c) day 42 (n = 8). d–g) The antibody titers of four hemagglutinin at day 82 (n = 8). A1, A3, BV, and BY represent influenza A virus H1N1, influenza A virus H3N2, influenza B virus Victoria, and influenza B virus Yamagata, respectively. 2*M‐CP represents double the dose of M‐CP adjuvant during immunizations. h,i) Proportion of the effector memory T cells (h, CD3 + CD44 + CD62L − ) and central memory T cells ((i) CD3 + CD44 + CD62L + ) in splenocytes after re‐stimulation with tetravalent influenza lysis at day 82 (n = 6). j) The OD value of IgG, IgG1, IgG2a, and IgG2b against the recombinant antigen glycoprotein E (gE) analyzed by ELISA (n = 8). k) ELISpot analysis of INF‐γ spot‐forming cells among 1 × 10 6 splenocytes after re‐stimulation with gE protein at day 28 (n = 4). The data were presented as mean ± SEM (n ≥ 3). The multi‐component comparisons data b–i) were analyzed by one‐way ANOVA with Turkey multiple comparisons post‐test. The two‐component data j,k) was analyzed by t‐test. ( * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The recombinant HBsAg protein, tetravalent influenza lysis antigens, recombinant antigen glycoprotein E (gE) of varicella‐zoster virus, and corresponding antibody detection ELISA kits were from Shenzhen Kangtai Biological Products Co., Ltd. (Shenzhen, China).

Techniques: Adjuvant, Vaccines, Injection, Virus, Lysis, Recombinant, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal:

Article Title: IL-18 contributes to the spontaneous development of atopic dermatitis-like inflammatory skin lesion independently of IgE/stat6 under specific pathogen-free conditions

doi: 10.1073/pnas.152337799

Figure Lengend Snippet: Spontaneous deviation of splenic CD4+ T cells into Th2 cells. (a) High serum levels of IgE and IgG1 in both types of transgenic mice. Sera were sampled from the various types of mice (36 weeks old) and their serum levels of various types of Ig were measured by ELISA. Data are represented as mean ± SD of triplicate cultures. Similar results were obtained in three independent experiments. (b and c) Spontaneous development of Th2 cells in both types of transgenic mice. Splenic CD4+ T cells from KCASP1Tg or WT littermates at 12 weeks of age (b) or from KIL-18Tg or WT littermates at 36 weeks of age (c) were isolated by membrane attack complexes and were incubated with immobilized anti-CD3 for 48 h. Concentration of IL-4 and IFN-γ in each supernatant was determined by ELISA. Data are represented as mean ± SD of triplicate cultures. Similar results were obtained in three independent experiments. (d) Increase of CD40L-expressing CD4+ T cells in both types of transgenic mice. Spleen cells were isolated from KCASP1Tg, KIL-18Tg, or WT littermate at 36 weeks of age, and their CD40L expression gated on CD4+ cells were identified by flow cytometry. A representative result is shown. Similar results were obtained in three independent experiments. ND, not detected.

Article Snippet: IL-4 and IFN-γ levels were measured by corresponding ELISA kits (Genzyme TECHNE).

Techniques: Transgenic Assay, Enzyme-linked Immunosorbent Assay, Isolation, Incubation, Concentration Assay, Expressing, Flow Cytometry