HEK 293T cells were transfected with the indicated N-terminally HA-tagged TNFR1-EYFP fusion protein constructs. TNFR1 expression was measured in EYFP-positive cells using anti-HA.11 (clone 16B12; Covance) and APC-polyclonal goat anti-mouse IgG secondary antibody (BD Pharmingen™). For total anti-HA.11 staining, cells were permeabilized with ice-cold 70% ethanol for 30 min before incubation with the primary antibody.
