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. 2017 Jan 20;3(1):e1601602. doi: 10.1126/sciadv.1601602

Fig. 1. ASXL1 associates with cohesin complex proteins.

Fig. 1

(A) Table of the most relevant proteins identified by LC-MS/MS in the affinity purification of ASXL1-associated proteins using FLAG-ASXL1 overexpressing HEK293T cells. Spectral counts (unique and total) for each interacting protein are shown. (B to E) Reciprocal IP and Western blotting confirmed interaction of ASXL1 with SMC1A, SMC3, and RAD21 in nuclear fraction derived from HEK293T cells transfected with pcDNA3.1+ (Vec) or FLAG-tagged ASXL1 (ASXL1). Nuclear extractions were subjected to IP using indicated antibodies against FLAG (B), SMC1A (C), SMC3 (D), or RAD21 (E). IB, immunoblot. (F) Western blot shows the endogenous interaction between ASXL1 and SMC1A, SMC3, and RAD21 in BM cells of WT mice. IgG, immunoglobulin G. (G) Gel filtration analysis of nuclear extracts from FLAG-ASXL1 overexpressing cells. ASXL1 and the cohesin complex were coeluted from a Superose 6 HR gel filtration column, as analyzed by Western blotting. The numbers over the lanes represent the eluted fraction numbers.