Figure 2
YBAP1 affects nuclear localization of YB-1. (a) Domain structure of YB-1 protein. (b) Schematic diagrams of wild-type, the 1–219 deletion mutant, and the point mutant PM1. The consensus amino acid sequences of RNP-1 in the CSD, which were mutated in PM1, are shown. (c,d) FLAG-YB-1 expression vectors pCMV-3xFLAG-YB-1 (WT, 1-219 and PM1) were transfected into HeLa cells with or without the YBAP1-myc expression vector pCI-neo-C1QBPmyc. Cells were immunostained with antibodies against FLAG (green) and c-myc (red) tags, and stained with DAPI. Nucleoli are indicated by white arrowheads (c). Localization of FLAG-YB-1 was examined for >100 cells and classified into four categories (N, signals restricted to the nuclei; N > C, nuclear signals stronger than cytoplasmic signals; N = C, equal intensity of cytoplasmic and nuclear signals; C, cytoplasmic signals stronger than nuclear signals). In cells co-transfected with YBAP1-myc expression plasmid, localization of FLAG-YB-1 in cells co-expressing YBAP1-myc was assessed. The quantitative results of three independent experiments are shown as relative cell number (%) in d. Bars, s.d.
