FIG. 7.

Transcriptional activity of mutant BSX1A in mammalian cells. (A) Schematic representation of myc-tagged full-length, truncated (ΔAD), and mutated (ΔNLS) BSX1A and a luciferase reporter construct. A putative NLS (₁₀₉RRRKAR₁₁₄) was mutated to ₁₀₉GRGKAR₁₁₄ (*). HD, homeodomain. (B) Luciferase activities of myc-tagged protein-transfected Hs683 cells; 0.2 μg of reporter plasmid was used for each transfection. All luciferase activities were normalized for transfection efficiency by measuring the Renilla luciferase activity of cotransfected pRL-SV40 vector (0.05 μg). The error bars indicate standard deviations. (C) Western analysis of myc-tagged wild-type and mutant BSX1A and BSX1B. The expression vectors were transfected 293T cells, and cell lysates were blotted with α-myc antibody (Sigma). (D) EMSA. GST-BSX1AΔAD, but not GST-BSX1AΔNLS, retains the DNA binding ability to BBS. (Below) Coomassie-stained GST fusion proteins used for EMSA.