Recruitment of multiple repressor proteins to the γ-promoter is dependent on PRMT5. (A) Western blot analysis of proteins immunoprecipitated from K562 cells expressing PRMT5-f with α-FLAG antibodies (Abs). A mock immunoprecipitation with normal rabbit immunoglobulin G (IgG) was used as a negative control. Abs used are indicated on the right. (B) Western blot analysis of extract from K562 cells expressing PRMT5-f fractionated by Superose 12 gel filtration. Column fractions were concentrated and analyzed using the antibodies indicated. (C) ChIP analysis on chromatin derived from PRMT5-f–expressing K562 cells with the stated Abs. (D) ChIP-ReChIP analysis on chromatin derived from FLAG immunoprecipitates from PRMT5-f K562 cells with the stated Abs. The precipitated DNA was amplified with primers specific for the γ-promoters. Enrichment was calculated relative to normal rabbit IgG. The error bars correspond to the SD. Each experiment was performed twice independently.