α cd16 cd32 fc block 2 4g2

For flow cytometric analysis, cell suspensions were pretreated with α-CD16/CD32 Fc-Block (2,4G2; BD Biosciences) and stained by standard procedures. Dead cells were excluded by staining with Fixable Viability Dye eFluor 780 (eBioscience). Cells were stained using the antibodies listed in Table S1.
PtC-reactive cells were identified by staining with PtC-liposome fluorescein-DHPE (FormuMax Scientific). Biotin was detected by using Streptavidin Qdot 605 (Molecular Probes/Invitrogen). Unconjugated α-Pten- and α-Foxo1 antibodies were detected by using α-rabbit IgG (H+L) F(ab′)₂ fragment-Alexa Fluor 647 (Cell Signaling). Intracellular flow cytometric staining was performed using the kits listed in Table S2 according to the manufacturer’s instructions.
Cells were acquired at a FACSCantoII flow cytometer (BD Biosciences). If not indicated otherwise, numbers in the dot plots indicate the percentages of cells in the respective gates, numbers in the histograms indicate the mean fluorescence intensity (MFI).

For flow cytometric analysis, cell suspensions were pre-treated with α-CD16/CD32 Fc Block (2,4G2; BD Bio-sciences). Dead cells were excluded by staining with Fixable Viability Dye eFluor 450 (eBioscience). Intracellular (IC) flow cytometry staining was performed using the ADG Fix&Perm Kit (Dianova). The detailed IC and extracellular staining procedure including the respective antibodies is provided in the Online Supplementary Appendix. Cells were acquired at a FACS Canto II flow cytometer (BD Bio-sciences). Analysis was performed using the FlowJo software (Tree Star).