Herein, we used MHC class II tetramers and Foxp3 gfp knockin mice to track the kinetics and magnitude whereby pathogen-specific Foxp3 + CD4 + and Foxp3 CD4 + cells are primed and expand after acute infection with recombinant Listeria monocytogenes (Lm) expressing the non-"self"-Ag 2W1S ...
GFP-positive Treg cells were sorted by FACS. The shRNA sequences were used as follow: shCYP1A1-1, 5′-GCTGACTTCATCCCTATTCTT-3′; shCYP1A1-2, 5′- GCACAACAAGAGACACAAGTT-3′. 2.4. Single-cell separation, library construction and RNA-seq We collected in vitro cultured human Tregs ...
(A) Naïve CD4 T cells isolated from C57BL/6J mice were cultured for 1 day under Th0 conditions and then transduced with the shRNA-Irs1 vector. Cells were then cultured for 2 days under suboptimal Treg-inducing conditions. GFP+ cells were used for transduction. FOXP3 expression was ...
Cell death in HT22 cells was strongly inhibited by a factor of approximately 2 after treatment with CM from cultures exposed to IL-4, CD200, or Foxp3-GFP, compared with pre-treatment of HT22 cells with LPS alone (Fig. 6B). As Foxp3-positive Treg cells have been found to induce ...
TM the day of the transfer. In vitro assays FACS-sorted CD4+Foxp3+ TREG cells (GFP+, 50 × 103) expressing ST2 (ST2+) or not (ST2-) were activated in 96-well flat-bottomed (0.2 ml) plates previously coated with α-CD3 (5 μg/ml) and α-CD28 (2 μg/ml), and...
(3.5 ml ml À 1 Dynabeads Mouse T-Activator CD3/CD28, 10 ng ml À 1 IL-23, 30 ng ml À 1 IL-6 and 2 ng ml À 1 TGF-b) medium for (2 72 h at ng ml À 37 °C. Half of the medium 1 TGF-b) for additional 24 was exchanged with h (Foxp3GFP subsets) Treg-...
B6.129(Cg)-Foxp3tm3(DTR/GFP)Ayr/J (B6 background) Jackson Laboratory JAX: 016958 B6.Cg-Nr3c1tm1.1Jda/J (B6 background) Jackson Laboratory JAX: 021021 B6.129(Cg)-Foxp3tm4(YFP/icre)Ayr/J (B6 background) Jackson Laboratory JAX: 016959 Oligonucleotides GAPDH forward:5’- ATGCCTGCTTCACCA...
(B) In vitro suppression assays with CXCR5+ and CXCR5− 4PD1hi, 4PD1− and Tregs from spleens (SP) and tumors (TM) of naive and B16-bearing (TB) Foxp3-GFP mice immunized with sRBC. Mean ± SD proliferation (CTVlow) of target CD45.1+CD4+ T cells and quantification of IL-2 by...
C57BL/6-Tg(Foxp3-DTR/EGFP)23.2Spar/Mmjax (DTR-Foxp3-GFP) The Jackson Laboratory Stock No: 032050 B6.129(Cg)-Foxp3tm4(YFP/icre)Ayr/J (Foxp3-YFP-Cre) (Foxp3-YFP-Cre) The Jackson Laboratory Stock No. 016959 C57BL/6-Tg(Foxp3-GFP)90Pkraj/J (Foxp3-GFP) The Jackson Laboratory Sto...