Figure 1. Identification of candidate interacting partners of LEDGF/p75 using transcription factor protein arrays.
(A) Coomassie blue stained SDS-PAGE gel showing purified His-LEDGF/p75. E. coli BL21 strain was transformed with pET28a-dfs70 encoding His-LEDGF/p75 and induced with IPTG. Lysate was passed through a nickel column to purify His-LEDGF/p75. (B) Immunoblot showing the specificity of the human autoantibody against LEDGF/p75 used as detection reagent in the transcription factor protein arrays. The autoantibody reacts specifically with LEDGF/p75 in a PC3 PCa cell lysate. (C) Transcription factor arrays were used to identify candidate interacting transcription factors of LEDGF/p75. Purified His-LEDGF/p75 was incubated with transcription factors spotted on membranes. Protein interactions were detected with human anti-LEDGF/p75 autoantibody and chemiluminescence. A section of the transcription factor array membrane containing MeCP2 is showed.