Partners of different NHSL3 isoforms

MCF10A cells were genome edited to tag the endogenous ORF of NHSL3 and generate C-terminal GFP-Flag fusion proteins of all NHSL3 isoforms. MCF10A cells express 4 NHSL3 isoforms through the use of alternative exons (i1, i2, i3L and i3S). Plasmids encoding N-terminal Flag-GFP fusion proteins were constructed and used to select stable MCF10A cell lines expressing each of the tagged isoform. The 4 distinct NHSL3 isoforms in these stable cell lines as well as all isoforms together from the genome edited cell line were purified by Tandem Affinity Purification and associated proteins were identified by LC-MS/MS.