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

Figure 3. The rCTF is toxic to cells when expressed in the cytoplasm.

Figure 3

(A) Cell death in transiently over-expressed HEK293T cells assessed by LDH level at 72 hours after transfection. The rCTF-polyQ-NES exhibits stronger cytotoxicity than rCTF-polyQ-NLS. (B) Time course of cell death in inducible rCTF-Q28-NES PC12 cell line. Cell toxicity is detected from Day5 and become prominent in Day6. (C) The cell death at Day6 compared in 6 inducible PC12 cell lines (rCTF-Q13, rCTF-Q13-NLS, rCTF-Q13-NES, rCTF-Q28, rCTF-Q28-NLS, rCTF-Q28-NES). The cell line expressing rCTF-Q28-NES exerts the strongest cell death. Y-axis shows differences of LDH values between (Dox(−)) and (Dox(+)) states. Cells do not show obvious cell death in Dox(+) states. (D&E) TUNEL positive cells are dramatically increased when doxycyclin is removed (Dox(−)). Eight randomly selected microscope fields were counted for TUNEL positive cells and total cell numbers. (For A to C and E: *: p<0.05; **: p<0.01; ***:p<0.001;ANOVA for A to C. Mann-Whitney's U test for E. Error bars indicate ± SEM.).