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 ...
(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...
Figure 4.IRS1 downregulates CD25 and CTLA4 on Treg cells and suppresses their immunosuppressive functions. IRS1 overexpression was performed as inFigure 2. (A) GFP+ Treg cells (transduced by MIEG3-IRS1) were gated, and expression of ICOS, GITR, CD25, and CTLA was measured via flow cytometry...
To further confirm Foxp3 expression in microglia, we utilized B6.Cg-Foxp3tm2Tch/J (Foxp3EGFP) mice, which co-express Enhanced Green Fluorescent Protein (EGFP) and Foxp3 under the control of its endog- enous promoter17. Weakly labelled EGFP-positive cells were detected in the hippocampi of...
(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-...
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...
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...
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...