ILDR1 is a novel PLSCR1-binding protein
In our previous study, we performed yeast two-hybrid screening of a
chicken cDNA library using the C-terminal intracellular domain of ILDR1
(228–553 aa) as bait (Liu et al., 2017). Among the positive clones
identified, five clones encode PLSCR1. We then performed
co-immunoprecipitation (co-IP) experiments to confirm the interaction
between ILDR1 and PLSCR1. The results show that EGFP-tagged mouse ILDR1
cytoplasmic domain could be co-immunoprecipitated together with
Myc-tagged PLSCR1 (Fig. 4A). Likewise, EGFP-tagged PLSCR1 could be
co-immunoprecipitated together with Myc-tagged ILDR1 cytoplasmic domain
(Fig. 4B). ILDR1-EGFP mainly localizes in the cytoplasm in COS-7 cells
as reported previously (Fig. 4C) (Hauge et al., 2004). In contrast,
PLSCR1-mCherry localizes in both cytoplasm and nucleus (Fig. 4D). When
cotransfected, ILDR1-EGFP translocates from cytoplasm to nuclei with
PLSCR1-Mcherry (Fig. 4E). Our co-IP and co-localization results suggest
that ILDR1 is a novel PLSCR1-binding partner.