Flow cytometry
Circulating Th2 cells were assessed as the proportion of peripheral
blood CD4+ T cells expressing CRTh2 as we described
previously [26,
34] (Fig. S1). CRTh2 levels on in
vitro differentiated Th2 cells were determined as previously referenced
[35] (Fig. S2). Briefly, cells were
stained using antibodies against CD4, CRTh2 or isotype
control
and fixed with paraformaldehyde (2%). Flow cytometry analysis was
performed using a BD LSR II Flow Cytometer, with gates set in accordance
with the profiles of the isotype control and/or negative control beads.
Results were analyzed using FlowJo® (TreeStar, Ashland
OR, USA).