Figure 3.
Subcellular localization of native or overexpressed mThrRS and mThrRS-L. Cytoplasmic (C) and nuclear (N) fractions were separated from NIH/3T3 cells or NIH/3T3 cells overexpressing mThrRS-L-FLAG or mThrRS-FLAG. The localization of native mThrRS-L in NIH/3T3 or GC2 (A), overexpressed mThrRS-L-FLAG, or mThrRS-FLAG in NIH/3T3 (B) was detected using anti-mThrRS-L or anti-FLAG antibodies. Note that a qualified anti-mThrRS antibody was not obtained, thus mGlyRS was detected using anti-mGlyRS in (A) as a comparison. α-Tubulin and Histone 3.1 were used as indicators of the cytoplasmic and nuclear fractions, respectively. The distribution pattern of α-Tubulin and Histone 3.1 between NIH/3T3 cells overexpressing mThrRS-L-FLAG or mThrRS-FLAG is the same; thus, only the localization of α-Tubulin and Histone 3.1 from NIH/3T3 cells overexpressing mThrRS-L-FLAG is shown in (B). The gene encoding mThrRS-L-EGFP, mThrRS-EGFP, mThrRS-L-Δ149-EGFP or mThrRS-L-A4-EGFP was overexpressed in HEK293T cells and the localization of each protein was observed. The nucleus was stained by DAPI (C).