Dissection of protein and RNA regions required for SPEN binding to XIST A-repeat RNA

XIST_A-full and SRA RNA bind SPEN_{RRM 1–4} with similar affinity. (A) Constructs of SPEN_{RRM 1–4}, the XIST A-repeat region (XIST_A-full), and SRA RNA used for binding studies. (B) EMSA gel-shift assay of XIST_A-full and SPEN_{RRM 1–4} interaction. (C) Quantification of bound fraction of XIST_A-full and SPEN_{RRM 1–4} with increasing protein concentration, n = 6. Data are the mean ± SEM. (D) EMSA gel-shift assay of SRA and SPEN_{RRM 1–4}. (E) Quantification of bound fractions of SRA and SPEN_{RRM 1–4}, n = 6. Data are the mean ± SEM. (F) Binding affinity of XIST_A-full and SRA RNA for SPEN_{RRM 1–4} protein construct, n = 6. (n.s.) Not significant, P > 0.05. Data are the mean ± SEM. (G) Competitive gel-shift assay between XIST_A-full (labeled) and SRA or BARD1 (unlabeled competitor) for binding to SPEN_{RRM 1–4}.