Figure 4.

Detection of Nitr9 isoforms by flow cytometry. HEK293T cells were transfected with plasmids encoding FLAG-tagged (a, c, and e) or endogenous (b, d, and f) isoforms of Nitr9 and EGFP as indicated above the panels. Cells were labeled with an anti-FLAG antibody (top row), anti-Nitr9⁹⁰ (middle row) or anti-Nitr9¹⁹ (bottom row), and an APC conjugated secondary antibody. Flow cytometric analyses were employed to detect EGFP positive (X axis) and APC positive (Y axis) cells. Isotype-matched antibodies were evaluated as controls for both anti-Nitr9 antibodies and displayed no labeling of transfected cells (data not shown).