Anti akt ps473 m89 61

Cell surface staining was performed using standard methods with the following antibodies: anti-CD4 (GK1.5, Biolegend, San Diego, CA), anti-CD8 (53-6.7, BD Pharmingen, San Jose, CA), anti-B220 (RA3-6B2, BD Pharmingen), anti-CD11b (M1/70, BD Pharmingen), anti-CD11c HL3, (BD Pharmingen) and CD25 (PC61, BD Pharmingen). Staining for FoxP3 was performed using the FoxP3 buffer set (eBioscience, San Diego, CA) according to the manufacturers instructions, anti-FoxP3 antibody (FJK16A, eBioscience), anti-Bim antibody (C34C5, Cell Signaling Technologies, Beverly, MA) and anti-AKT pS473 (M89-61, BD Pharmingen). Central (cTregs) and effector Tregs (eTregs) were analyzed as described in reference (54 (link)). All flow cytometry were performed on a FacsCanto II cytometer (BD Biosciences, San Jose, CA), and data were analyzed using FlowJo v X 10.0.7 software (Tree Star, Inc., Ashland, OR).

Cell surface staining was performed using standard methods. The following antibodies were used: Anti-CD4 (GK1.5, Biolegend, San Diego, CA), anti-CD3 (2C11, Biolegend), anti-CD8α (53–6.7, Biolegend), anti-FoxP3 antibody (FJK16A, eBioscience), anti-Akt pS473 (M89–61, BD Pharmingen), anti-human CD2 (hCD2- PE), anti-CD44 (IM7, Biolegend), anti-CD62L (MeL14, Biolegend), anti-Pten (BD Pharmingen), anti-PD1 (J43, BD Pharmingen).