Supplemental Fig 1. Representative FC plot gating strategy to
identify live CD45+ CD3+ CD4+ CXCR1+ IL4+ Th2-cells in the lungs
The forward and side scatter (FSC vs SSC) density plots were generated
to separate cell populations of interest from debris. Next, doublets and
debris were excluded by forward scatter height versus forward scatter
area density plot (FSC-H vs FSC-A). Next, dead cells were excluded by
gating only live cells using fluorophore-conjugated antibodies Fixable
Viability Dye eFluor 780. In all subsequent steps, side scatter area
(SSC-A) was plotted against fluorescence-tagged antibodies to gate
positive cells by excluding cells in the corresponding FMO gate.
Sequentially, live cells were gated using CD45+ cells to gate
hematopoietic cells, CD3+, CD4+ double positive cells to gate for CD4+
T-cells, CXCR1+ cells to gate CXCR1-expressing CD4+ T-cells, then
intracellular IL4+ to gate for CXCR1+ IL4+ Th2-cells in the lungs.
FMO; fluorescence minus one