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. 2013 Mar 7;8(3):e50121. doi: 10.1371/journal.pone.0050121

Figure 1. Localization signals efficiently change the intracellular location of CTF.

Figure 1

(A) Recombinant Cav2.1 C-terminal fragment (rCTF)s used in this study. Definition of rCTF (amino acid(AA) #1954–2506 in Cav2.1 [GenBank AB035726] [18]), artificial nuclear localization or export signals (NLS or NES), two different polyglutamine (Qn) (Q13; Q28) and recognition sites of three different antibodies (1C2 against expanded polyQ, A6RPT-#5803 against Cav2.1 CTF, c-Myc antibody against Myc-tag) are shown. (B) The rCTF-Q13 is predominantly expressed in the cytoplasm of PC12 cells. The NLS and NES efficiently localize the tagged rCTF to the designed location [A6RPT-#5803 antibody (red), Hoechst 33258 (blue)] (scale bar: 10 µm). (C) The NLS and NES efficiently shift rCTFs with expanded polyQ (Q28) into the nucleus and cytoplasm respectively (scale bars: 10 µm). (D) The proportion of the subcellular localization in each rCTF expressed. The rCTF-polyQ is predominantly, though not exclusively, expressed in the cytoplasm of PC12 cells. The localization signals change subcellular localization of rCTFs-polyQ effectively. (N; the cells expressing rCTF exclusively in the nucleus; N-c: the cells expressing rCTF predominantly in the nucleus than in the cytoplasm; n-C: the cells expressing rCTF predominantly in the cytoplasm than in the nucleus; C: the cells expressing rCTF exclusively in the cytoplasm) (For D: ***:p<0.001;ANOVA. Error bars indicate ± SEM.).