Purification and characterization of the SFPQ/NONO complex. (A) Enrichment of the SFPQ/NONO complex from HeLa nuclear cell extracts using Ni-NTA PhyCap OTC including the absence of Ni2+ and NTA. (B) Ni-NTA PhyCap OTC enrichment of the SFPQ/NONO complex from different cell extracts. Purified complexes were analyzed on SDS-PAGE stained with Coomassie blue. (C) Functional categorization of proteins identified using mass spectrometry analysis of the SFPQ/NONO complex purified using Ni-NTA PhyCap OTCs. (D) Effect of salt concentration on the SFPQ/NONO interaction. Ni-NTA PhyCap OTC enrichment of the SFPQ/NONO complex was performed over a range of NaCl concentrations highlighted. (E) Effect of splicing conditions on the SFPQ/NONO interaction. Ni-NTA PhyCap OTC enrichment of the SFPQ/NONO complex was performed under both splicing and nonsplicing conditions as shown. (F) Repurification of the SFPQ/NONO complex under low salt conditions (6 mM NaCl). (G) Chemical crosslinking of the SFPQ/NONO complex. Following chemical crosslinking the complex was analyzed using SDS-PAGE stained with Coomassie blue.
