Fig. 3.

NCDN interacts with SNRPN, SNRPB and SMN in cell lines and in mice. (A) Affinity isolation of NCDN–GFP using GFP-Trap, detected with anti-GFP antibody (top row) co-enriches mCherry–SNRPB, detected with anti-mCherry (bottom row) in transiently co-transfected SH-SY5Y cells. (B) In an SH-SY5Y cell line constitutively expressing NCDN–GFP, affinity isolation of NCDN–GFP, detected with anti-GFP antibody (top row) co-enriches SMN, SNRPB, SNRPN and the coatomer protein βCOP, all detected with antibodies against the endogenous proteins (as labelled). (C) In an SH-SY5Y cell line constitutively expressing YFP, affinity isolation of YFP, detected with anti-GFP antibody (top row) does not co-enrich SMN, SNRPB or βCOP, all detected with antibodies to the endogenous proteins (as labelled). (D) In an SH-SY5Y cell line constitutively expressing GFP–SMN, affinity isolation of GFP–SMN, detected with anti-GFP antibody (top row) co-enriches endogenous NCDN, detected with anti-NCDN antibody (middle row). Endogenous SMN, detected with anti-SMN (bottom row) is also co-enriched. (E) Immunoprecipitation (IP) of endogenous SMN co-enriches endogenous NCDN in SH-SY5Y cells. (F) Immunoprecipitation of endogenous SMN from murine P8 brain lysate co-enriches NCDN.