rat il-6 elisa kit  (Multi Sciences (Lianke) Biotech Co Ltd)

Journal: eLife

Article Title: An IL-21R STAT hypomorph circumvents functional redundancy in germinal centre responses

doi: 10.7554/eLife.109834

Figure Lengend Snippet: ( A ) Th17 cell conversion efficiency was compared using splenic CD4 + T cells from WT and Il21r EINS mice assessed on day 3 of stimulation after 4 hr of restimulation with PMA and ionomycin. IL-17 containing CD4 + T cells were assessed by intracellular immunostaining and flow cytometry. IL-6 signalling was blocked in vivo by the injection of 0.5 mg neutralizing anti-IL-6 mAb or isotype control mAb on day 0 followed by 0.25 mg on days 2, 4, and 6. Splenocytes from WT, Il21r −/− , and Il21r EINS mice were stimulated with rmIL-6 for 15 min at the concentrations shown and phosphorylated STAT1 ( B ) and phosphorylated STAT3 ( C ) detected in CD4 + T cells by intracellular immunostaining and flow cytometry. Mice were immunized with sheep red blood cell (SRBC) on day 0 and analysed on day 7 by flow cytometry and FACS analyses. Shown here are the percentages of ( D ) Tfh cells within the CD4 + T cells population and ( E ) the germinal centre (GC) B cell population as a percentage of total B cells. Data are shown as individual mice with n = 5 per group from 2 separate experiments with similar results. Statistical significance was assessed by ( A–C ) one-way ANOVA using Bonferroni’s multiple comparisons test and Student’s t -test ( D, E ); *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: For in vivo neutralization of IL-6: Mice initially received 0.5 mg neutralizing anti-IL-6 (clone MP5-20F3, rat IgG1; BioXCell) or isotype mAb (anti-rat IgG1; BioXCell), and subsequently given 0.25 mg mAb (i.v.) every other day following the initial dose through the end of the experiment ( ).

Techniques: Immunostaining, Flow Cytometry, In Vivo, Injection, Control

Journal: eLife

Article Title: An IL-21R STAT hypomorph circumvents functional redundancy in germinal centre responses

doi: 10.7554/eLife.109834

Figure Lengend Snippet: Analysis of the GC response in the spleen in WT, Il21r −/− , and Il21r EINS mice on day 7 following sheep red blood cell (SRBC) immunization. IL-6 receptor (IL-6Rα) expression on CXCR5 hi PD1 hi FoxP3 − T follicular helper (Tfh) cells from the spleen of WT, Il21r −/− , and Il21r EINS mice showing percentage ( A ) and mean fluorescence intensity (MFI) ( B ) and quantitation of MFI of IL-6Rα on Tfh cells ( C ). IL-6 signalling was blocked in vivo by the injection of 0.5 mg neutralizing anti-IL-6 mAb or isotype mAb and 0.25 mg on day 0, the day of SRBC immunization, and then days 2, 4, and 6. GC populations were analysed in the spleen on day 7. ( D ) Percentage of Tfh cells of CD4 + T cells, ( E ) percent IgG1 + FAS + B cells of total B cells. Data shown as individual mice ( n = 4–5 per group) with 2 experimental repeats showing similar results. Statistical significance was assessed by one-way ANOVA using Bonferroni’s multiple comparisons test; * p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: For in vivo neutralization of IL-6: Mice initially received 0.5 mg neutralizing anti-IL-6 (clone MP5-20F3, rat IgG1; BioXCell) or isotype mAb (anti-rat IgG1; BioXCell), and subsequently given 0.25 mg mAb (i.v.) every other day following the initial dose through the end of the experiment ( ).

Techniques: Analysis, Expressing, Fluorescence, Quantitation Assay, In Vivo, Injection