Journal: Frontiers in Immunology

Article Title: The iNKT cell ligand α-GalCer prevents murine septic shock by inducing IL10-producing iNKT and B cells

doi: 10.3389/fimmu.2024.1457690

Figure Lengend Snippet: α-GalCer pretreatment alters iNKT cell subsets and attenuates sepsis severity. WT B6 mice were injected i.p. with α-GalCer (2 μg/mouse) and, seven days later, the spleens were harvested for the following analyses. (A) Weight and total cell number of spleens. (B) The frequency and cell number of splenic iNKT cells (α-GalCer/CD1d-dimer + CD3 + ). (C) The frequency of CD4 + iNKT cells in the spleen. (D) The relative frequencies of iNKT cell subsets (i.e., T-bet + iNKT1, PLZF + iNKT2, RORγt + iNKT17, and E4BP4 + iNKT10 cells) were determined by flow cytometry. (E) Experimental outline: WT B6 mice were injected i.p. with α-GalCer (2 μg/mouse) and, seven days later, these mice were injected i.p. with LPS (2 µg/mouse) plus D-GalN (25 mg/mouse) for induction of sepsis. (F) Subsequently, these mice were monitored to evaluate their survival for three days. (G, H) H&E staining of liver sections (CV, central vein) (G) and serum levels of AST and ALT (H) were analyzed five hours after LPS/D-GalN injection. The mean values ± SD ( n = 3 in (A–D, H) ; per group in the experiment; Student’s t -test; * p < 0.05, ** p < 0.01, and *** p < 0.001) are shown. The survival rate was analyzed by Kaplan-Meier plots with a log-rank test (* p < 0.05). One representative experiment of two experiments is shown. ns, not significant.

Article Snippet: The following mAbs from Thermo Fisher Scientific (Waltham, MA, USA) were used: FITC-, PE- or allophycocyanin (APC)-conjugated anti-CD19 (clone 1D3); FITC-conjugated anti-F4/80 (clone BM8); FITC-conjugated anti-NK1.1 (clone PK-136); PE-conjugated anti-FcϵRI (clone MAR-1); PE-conjugated anti-T-bet (clone 4B10); PE-conjugated anti-E4BP4 (clone S2M-E19); PE-conjugated anti-Nur77 (clone 12.14); PE-conjugated anti-PLZF (clone Mags.21F7); PE-conjugated anti-IL17A (clone eBio17B7); PE-conjugated anti-IL1β (clone NJTEN3); PE/Cy7-conjugated anti-CD23 (clone B3B4); PE/Cy7-conjugated anti-CD3ϵ (clone 145-2C11); FITC- or APC-conjugated anti-Ly6G (Gr-1) (clone 1A8-Ly6g); APC-conjugated anti-CD200R3 (clone Ba13).

Techniques: Injection, Flow Cytometry, Staining

Journal: Frontiers in Immunology

Article Title: The iNKT cell ligand α-GalCer prevents murine septic shock by inducing IL10-producing iNKT and B cells

doi: 10.3389/fimmu.2024.1457690

Figure Lengend Snippet: The preventive effects of α-GalCer pretreatment on sepsis do not correlate with the IL4-STAT6 signaling pathway. WT B6 mice were injected i.p. with α-GalCer (2 μg/mouse) or OCH (2 μg/mouse) and, seven days later, the spleens were harvested. (A) The frequency and absolute cell number of splenic iNKT cells were determined by flow cytometry. (B) Flow cytometric analysis for the expression of transcription factors (T-bet, PLZF, RORγt, and E4BP4) by splenic iNKT cells. (C) WT B6 mice were injected i.p. with α-GalCer (2 μg/mouse) or OCH (2 μg/mouse) and, seven days later, mice were injected i.p. with LPS (2 µg/mouse) plus D-GalN (25 mg/mouse) for induction of sepsis. Subsequently, these mice were monitored to evaluate their survival for three days after LPS/D-GalN injection. (D) Experimental outline: WT B6 mice were injected i.p. with α-GalCer (2 μg/mouse) on day 0 and, seven days later, mice were injected i.p. with LPS (2 µg/mouse) plus D-GalN (25 mg/mouse) for induction of sepsis. To evaluate the effect of IL4 signaling on α-GalCer-mediated attenuation of sepsis, WT B6 mice were injected i.p. four times with a STAT6 inhibitor (AS1517499, 10mg/kg) every other day starting from day 0. (E) Subsequently, these mice were monitored to evaluate their survival for three days after LPS/D-GalN injection. The mean values ± SD (n = 4 in (A, B) ; per group in the experiment; Student’s t -test; ** p < 0.01 and *** p < 0.001) are shown. The survival rate was analyzed by Kaplan-Meier plots with a log-rank test (** p < 0.01 and *** p < 0.001). One representative experiment of two experiments is shown. ns, not significant.

Article Snippet: The following mAbs from Thermo Fisher Scientific (Waltham, MA, USA) were used: FITC-, PE- or allophycocyanin (APC)-conjugated anti-CD19 (clone 1D3); FITC-conjugated anti-F4/80 (clone BM8); FITC-conjugated anti-NK1.1 (clone PK-136); PE-conjugated anti-FcϵRI (clone MAR-1); PE-conjugated anti-T-bet (clone 4B10); PE-conjugated anti-E4BP4 (clone S2M-E19); PE-conjugated anti-Nur77 (clone 12.14); PE-conjugated anti-PLZF (clone Mags.21F7); PE-conjugated anti-IL17A (clone eBio17B7); PE-conjugated anti-IL1β (clone NJTEN3); PE/Cy7-conjugated anti-CD23 (clone B3B4); PE/Cy7-conjugated anti-CD3ϵ (clone 145-2C11); FITC- or APC-conjugated anti-Ly6G (Gr-1) (clone 1A8-Ly6g); APC-conjugated anti-CD200R3 (clone Ba13).

Techniques: Injection, Flow Cytometry, Expressing

Journal: Cell reports

Article Title: G2 arrest primes hematopoietic stem cells for megakaryopoiesis

doi: 10.1016/j.celrep.2024.114388

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Fluorescent Mounting Media with DAPI , OriGene , E19-18.

Techniques: Recombinant, Adhesive, Purification, Blocking Assay, Amplification, Random Hexamer Labeling, Reverse Transcription, SYBR Green Assay, Sterility, Imaging, Software

Journal: Cell reports

Article Title: G2 arrest primes hematopoietic stem cells for megakaryopoiesis

doi: 10.1016/j.celrep.2024.114388

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Fluorescent Mounting Media with DAPI , OriGene , E19-18.

Techniques: Recombinant, Adhesive, Purification, Blocking Assay, Amplification, Random Hexamer Labeling, Reverse Transcription, SYBR Green Assay, Sterility, Imaging, Software