Anti cd127 a7r34

Flow cytometry antibodies for anti-CD127 (A7R34) and CD122 (2-1221-82) were purchased from eBioscience (San Diego, CA, USA): anti-CD3ε (BD Biosciences, San Diego, CA, USA), anti-CD8 (4B11) (Invitrogen, Carlsbad, CA, USA), anti-KLRG1 (2F1) (Southern Biotech, Birmingham, AL, USA), anti-TGFβRI, anti-Smad2/3, anti-Smad4 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-TGFβRII (R&D Systems), anti-T-bet (BioLegend, San Diego, CA, USA), and anti-RGS3. The anti-RGS3 antibody has been described previously [10 (link)]. DAPI was purchased from Invitrogen. Extracellular and intracellular marker staining was performed as previously described [11 (link)]. Flow cytometric analysis was performed using an LSRII flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA). Data were analyzed using Flow Jo software (Tree Star, Ashland, OR, USA). The cells were fixed and permeabilized with methanol and formaldehyde prior to staining for analysis with the Amnis Imagestream imaging flow cytometer. Data were analyzed using Amnis IDEAS software v4.0 (Amnis Corporation, Seattle, WA, USA).