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

Journal: Redox Biology

Article Title: CO-mediated restoration of mitochondrial redox homeostasis contributes to alleviating multiple programmed cell deaths in hepatic I/R injury

doi: 10.1016/j.redox.2026.104440

Figure Lengend Snippet: In vivo therapeutic evaluation of CPs@HA in HIRI. (A) Schematic illustration of the experimental timeline for the HIRI model (created with BioRender.com ). (B) In vivo fluorescence imaging of Cy5.5-CPs and Cy5.5-CPs@HA , showing liver accumulation at different time points after intravenous injection. (C) Quantitative analysis of liver fluorescence intensity at 0, 4, 8, 12, and 24 h post-injection. (D-F) Serum levels of ALT, AST, and BUN in HIRI mice. (G) Representative images of TUNEL and H&E staining of liver tissues from different treatment groups (scale bar = 50 μm and 100 μm). (H) Quantification of TUNEL-positive area. (I) IF staining for the oxidative DNA damage marker 8-OHdG, the M1-type macrophage marker CD86, and the M2-type macrophage marker CD206 in liver sections (scale bar = 100 μm). (J, K) Activities of GSH-Px and SOD in liver tissues. (L-O) Inflammatory cytokine expression levels (IL-1 β , IL-6, IL-4, and IL-10) in liver tissues determined by ELISA. (P) PCA of samples from the Normal, HIRI and CPs@HA groups. (Q) GSEA comparing the Normal and HIRI groups. (R) GSEA comparing the HIRI and CPs@HA groups. Statistical analyses were performed by comparing the HIRI group with each of the other groups ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 vs . indicated. ns, not significant.

Article Snippet: Antibodies against DLAT, G6PD, FLNA, p -AMPK α , AMPK α , SIRT1, PGC-1 α , CD86, HO-1, CD163, 8-OHdG, iNOS, and GPX4, as well as mouse IL-4, IL-10, and IL-6 ELISA kits, were purchased from Proteintech (USA).

Techniques: In Vivo, Fluorescence, Imaging, Injection, Analysis, TUNEL Assay, Staining, Marker, Expressing, Enzyme-linked Immunosorbent Assay, H&E Stain, RNA Sequencing