Team:Kyoto/CiC/Results

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Results & Discussion

Result of Subgoal A

Confirmation of function of TAT-liposome

To confirm the function of the constructed TAT-anchor peptide, we prepare the liposome containing NBD-PE with or without TAT-anchor which was synthesized or translated in vitro. The cells incubated with liposome did not emit any fluorescent signal.

Result of Subgoal B

Result of Subgoal C

sig-GFP expression in HeLa

To confirm the function of the signal sequence for importing protein into mitochondria, we compared the expressioin pattern of sig-GFP or GFP with mitotracker signal. The GFP signal was detected throughout the cell except for the black granules in the cytoplasm or the nuclear, while sig-GFP signal showed the string-like pattern in the cytoplasm. The black granules in the cytoplasm observed in the GFP-expressing cell were stained by mitotracker, and the result indicated that the GFP was normally eliminated from mitochondria (Fig. NUMBER, GFP and mitotracker merged). In case of sig-GFP, mitochondria stained by a mitotracker and the GFP signal showed almost the same pattern. the yellow color in the merge images (Fig. Number, sig-GFP and mitotracker merged) suggested that the sig-GFP and mitochondria were colocalized in the HeLa cell. We, consequently, our constructed signal sequence has the function of importing protein into mitochondria as expected.

 Figure Caption: Conforcal microscopic images of GFP or sig-GFP transfected cells. The image lines titled
 "mitotracker(+)" indicated the samples was stained by mitotracker. The columns showed the
 mitotracker, GFP, and merged images from left, respectively.