Fig. 1
NKp30 interacts with DPEP1. (A) Co-immunoprecipitation of NKp30 and DPEP1. After co-culturing NK92 and KM12C cells, cell lysates were immunoprecipitated (IP) using either anti-DPEP1 or anti-NKp30 antibodies. Immunocomplexes were analyzed by SDS–PAGE followed by western blotting. (B) Interaction of NKp30 isoforms with DPEP1. NK92 cells were transfected with His-tagged NKp30 isoforms (a/b/c), co-cultured with KM12C cells, and subjected to IP using anti-DPEP1 antibody. NS, not specific. (C) Binding of recombinant DPEP1 to NK92 cells. rDPEP1 was purified from HEK293F cells and visualized by Coomassie Brilliant Blue staining. NK92 cells were treated with rDPEP1, washed, and stained with anti-NKp30/APC and anti-DPEP1/FITC antibodies. NKp30⁺/rDPEP1⁺ NK92 cells were quantified by flow cytometry. Data represent mean ± SD from three independent experiments (n = 3), each performed in triplicate. Statistical significance was assessed using Student’s t-test (**P < 0.01). (D) Colocalization of rDPEP1 and NKp30 on NK92 cells. Cells treated with or without rDPEP1 were fixed and stained with anti-NKp30/FITC and anti-DPEP1/TRITC antibodies. Nuclei were counterstained with DAPI. Arrows indicate colocalization of both proteins on the plasma membrane (confocal microscopy).
