Journal: Journal of Virology

Article Title: NFAT4 Is Required for JC Virus Infection of Glial Cells

doi: 10.1128/jvi.01456-06

Figure Lengend Snippet: FIG. 2. NFAT4 is functionally active in glial cells. (a) NFAT expression in SVG-A and U-87MG whole-cell lysates. NFATs 1 to 4 were detected using antibodies (NFATc1 7A6, NFATc2 4G6-G5, NFATc3 F-1, and NFATc4 H-74; Santa Cruz Biotech Inc.) diluted 1:200. Ramos cell lysates were used as a positive control. (b) Luciferase reporter gene assays using an NFAT-responsive reporter construct were used to compare NFAT activity in SVG-A cells and U-87MG cells in the presence or absence of 292 mg/liter glutamate. A control construct that lacked the NFAT binding site was used to measure basal transcriptional activity. (c) The NFAT reporter construct was cotransfected with either a control construct ( SV40 T-Ag) or a construct expressing the SV40 large T-Ag ( SV40 T-Ag).

Article Snippet: Ramos cell lysates were purchased from Santa Cruz Biotech Inc. Whole-cell extracts were separated on Tris-HCl-ready gels (Bio-Rad).

Techniques: Expressing, Positive Control, Luciferase, Construct, Activity Assay, Control, Binding Assay

Journal: Journal of leukocyte biology

Article Title: Pivotal advance: alpha-galactosylceramide induces protection against lipopolysaccharide-induced shock.

doi: 10.1189/jlb.0506298

Figure Lengend Snippet: Fig. 3. Cytokine production by spleen cells from LPS-KRN7000-treated mice before challenge and reexposed in vitro to -GalCer. C57BL/6 mice were injected with KRN7000 or with vehicle at indicated times before LPS challenge. 2 h after LPS challenge, groups of 10 mice were sacrificed. Pooled spleen cells from each group were cultured in complete medium with 10 ng/ml of KRN7000 (f) for 60 h at 37°C in 5% CO2 atmosphere. IFN (A), IL-4 (B), and IL-10 (C) were measured in culture supernatants by ELISA. Open triangles ( ) represent the levels of cytokines in mice treated with LPS-KRN7000 at the same times of previously showed experimental groups treated before challenge, and spleen cells were reexposed in vitro to vehicle. (A) Open circles () represent the levels of cytokines produced by splenocytes of mice treated with LPS and vehicle and reexposed in vitro to -GalCer. Results reported are the means of 3 different experiments. **, P 0.001.

Article Snippet: For measurement of cytokines levels, either in serum samples or in culture supernatants, we used IFN- , IL-4, IL-10, and TNF- Quantikine Mouse ELISA kits (R&D Systems, Minneapolis, MN, USA).

Techniques: In Vitro, Injection, Cell Culture, Enzyme-linked Immunosorbent Assay, Produced

Journal: Journal of leukocyte biology

Article Title: Pivotal advance: alpha-galactosylceramide induces protection against lipopolysaccharide-induced shock.

doi: 10.1189/jlb.0506298

Figure Lengend Snippet: Fig. 2. Cytokines levels in sera of mice treated with KRN7000 before challenge. Groups of 10 mice were injected with KRN7000 or vehicle only from 1 to 18 h before LPS challenge. Sera were collected from 10 mice of each group 1 h after challenge for TNF- (A), 6 h after challenge for IFN- (B), and 2 h after challenge for IL-10 assay (C) (f). Data reported are the means of serum cytokines content tested for each animal of any group. Differences from cytokines levels detected in controls and experimental groups were reproduced without significant modifications when tested sera were collected at different time points following LPS challenge (data not shown). Serum cytokine levels were determined by ELISA. Data are the means of three different experiments. Before LPS challenge, the above cytokines were undetectable in the sera. Open triangles ( ) represent the values obtained from mice treated with LPS plus vehicle at the same times of experimental groups treated before challenge. In these control groups, sera were collected after challenge at the same times of experimental groups. **, P 0.001 *, P 0.05.

Article Snippet: For measurement of cytokines levels, either in serum samples or in culture supernatants, we used IFN- , IL-4, IL-10, and TNF- Quantikine Mouse ELISA kits (R&D Systems, Minneapolis, MN, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of leukocyte biology

Article Title: Pivotal advance: alpha-galactosylceramide induces protection against lipopolysaccharide-induced shock.

doi: 10.1189/jlb.0506298

Figure Lengend Snippet: Fig. 4. KRN7000’s effects on systemic release of cytokines by mice challenged with LPS. Mice challenged with LPS were injected with KRN7000 or vehicle at indicated times after challenge. Sera of 10 mice for each experimental group were collected 4 h for TNF- (B), 6 h for IFN- (A), and 4 h for IL-10 (C) after challenge (f). Data reported are the means of serum cytokine content tested for each animal of any group. Open triangles ( ) represent the values obtained from sera of mice treated with LPS plus vehicle at the same times of experimental groups. Serum cytokine levels were determined by ELISA. Data represented are the means of 3 different experiments. Before LPS challenge, the above cytokines were undetectable in the sera. Differences from cytokine levels detected in controls and experimental groups were reproduced without significant modifications when sera tested were collected at different time points after LPS challenge (data not shown). **, P 0.001, *, P 0.05.

Article Snippet: For measurement of cytokines levels, either in serum samples or in culture supernatants, we used IFN- , IL-4, IL-10, and TNF- Quantikine Mouse ELISA kits (R&D Systems, Minneapolis, MN, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal: Journal of leukocyte biology

Article Title: Pivotal advance: alpha-galactosylceramide induces protection against lipopolysaccharide-induced shock.

doi: 10.1189/jlb.0506298

Figure Lengend Snippet: Fig. 5. KRN7000 in vitro reexposure induces poor IFN but high IL-4 and IL-10 production by spleen cells from LPS challenged-GalCer-protected mice. C57BL/6 mice were injected at indicated times after LPS challenge with KRN7000 or with vehicle. One hour after -GalCer administration, groups of 10 mice were sacrificed. Pooled spleen cells from each group were cultured in complete medium with 10 ng/ml of KRN7000 (f) or vehicle ( ) for 60 h at 37°C in 5% CO2 atmosphere. IFN- (A), IL-4 (B) and IL-10 (C) were measured in cultured supernatants by ELISA assays. Open triangles represent cytokine levels in mice primed, challenged with LPS, treated with KRN7000 and reexposed in vitro to vehicle. (A) Open circles () represent the levels of cytokines produced by splenocytes of mice primed, challenged with LPS plus vehicle, and reexposed in vitro to -GalCer. Results reported are the means of 3 different experiments. **, P 0.001.

Article Snippet: For measurement of cytokines levels, either in serum samples or in culture supernatants, we used IFN- , IL-4, IL-10, and TNF- Quantikine Mouse ELISA kits (R&D Systems, Minneapolis, MN, USA).

Techniques: In Vitro, Injection, Cell Culture, Enzyme-linked Immunosorbent Assay, Produced

Journal: Journal of leukocyte biology

Article Title: Pivotal advance: alpha-galactosylceramide induces protection against lipopolysaccharide-induced shock.

doi: 10.1189/jlb.0506298

Figure Lengend Snippet: Fig. 6. Cytokine production by NKT-negative and NK-negative fractions of protected and nonprotected mice. Spleen cells from KRN7000-protected (A) or nonprotected mice (B) were depleted of NKT and/or NK cells by immunomagnetic methods. NKT- and NK- fractions were obtained from protected or nonprotected mice. 4 105 of separated or unseparated cells were reexposed in vitro to -GalCer or vehicle dissolved in complete medium for 60 h at 37°C in 5% CO2 atmosphere. Cultured supernatants were tested for cytokines content by Quantikine ELISA kits. No significant difference in cytokines levels was detected testing both protected (treated with KRN7000 1.5 h before challenge or 1 h after challenge) or nonprotected groups (treated 18 h before challenge or 3 h after challenge). Data shown are the means of 3 different experiments.

Article Snippet: For measurement of cytokines levels, either in serum samples or in culture supernatants, we used IFN- , IL-4, IL-10, and TNF- Quantikine Mouse ELISA kits (R&D Systems, Minneapolis, MN, USA).

Techniques: In Vitro, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Device

Article Title: CRISPR-Cas12a immunosensing on glass fiber for point-of-care quantification of multiple inflammation biomarkers in osteoarthritis

doi: 10.1016/j.device.2024.100319

Figure Lengend Snippet: Figure 6. Detection of inflammation biomarkers in clinical samples of OA patients by the GUSCI platform and ELISA (A) Heatmap of the levels of protein expression. F indicates GUSCI platform results, while E indicates ELISA results. The color scale bar represents the con- centration of biomarkers (the concentration unit for IL-4, IL-6, TNF-a, IFN-g is pg mL1, and the unit for MMP-3 is ng mL1). (B–F) Correlation between the GUSCI platform and ELISA to measure IL-4, IL-6, TNF-a, IFN-g, and MMP-3 concentration.

Article Snippet: The human IL-6 ELISA kit (catalog number VAL102), human IL-4 ELISA kit (catalog number VAL123), and human IFN-g ELISA kit (catalog number VAL104C) were obtained from R&D Systems.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Concentration Assay

Journal: Vaccines

Article Title: An Antibody Persistent and Protective Two rSsCLP-Based Subunit Cocktail Vaccine against Sarcoptes scabiei in a Rabbit Model

doi: 10.3390/vaccines8010129

Figure Lengend Snippet: Variation of specific IgG ( a and b ) and total IgE ( c ) levels in the sera of immunized rabbits. a: the OD 450 nm value of the specific IgG antibodies was detected by an rSsCLP5-based indirect ELISA; b: the OD 450 nm value of specific IgG antibodies was detected by an rSsCLP12-based indirect ELISA; c: the total IgE antibody concentration (pg/mL) was detected by an ELISA kit (Elabscience, Wuhan, China). The rSsCLP 5 and rSsCLP 12 refer to rSsCLP5 and rSsCLP12, respectively.

Article Snippet: The serum samples collected from the vaccination trial were analyzed for the presence of specific IgG antibodies with a rSsCLP5-based indirect ELISA [ ] and rSsCLP12-based indirect ELISA [ ], total IgE antibodies with an ELISA kit (Elabscience, Wuhan, China), and the level of IL-4, IL-10, IFN-γ, and TNF-α with an ELISA kit (Elabscience, Wuhan, China).

Techniques: Indirect ELISA, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Vaccines

Article Title: An Antibody Persistent and Protective Two rSsCLP-Based Subunit Cocktail Vaccine against Sarcoptes scabiei in a Rabbit Model

doi: 10.3390/vaccines8010129

Figure Lengend Snippet: Variation of cytokines IL-10 ( a ), TNF-α ( b ), IL-4 ( c ), and IFN-γ ( d ) in the sera of immunized rabbits. a, the concentration of IL-10 (pg/mL) was detected by an ELISA kit (Elabscience, Wuhan, China); b, the concentration of TNF-α (pg/mL) was detected with an ELISA kit (Elabscience, Wuhan, China); c, the concentration of IL-4 (pg/mL) was detected using an ELISA kit (Elabscience, Wuhan, China); d, the concentration of IFN-γ (pg/mL) was detected using an ELISA kit (Elabscience, Wuhan, China). The rSsCLP 5 and rSsCLP 12 refer to rSsCLP5 and rSsCLP12, respectively.

Article Snippet: The serum samples collected from the vaccination trial were analyzed for the presence of specific IgG antibodies with a rSsCLP5-based indirect ELISA [ ] and rSsCLP12-based indirect ELISA [ ], total IgE antibodies with an ELISA kit (Elabscience, Wuhan, China), and the level of IL-4, IL-10, IFN-γ, and TNF-α with an ELISA kit (Elabscience, Wuhan, China).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay