FIGURE 5.

Dose-dependent TAT-mediated wt α-synuclein delivery and effects on mitochondrial Ca²⁺ transients and autophagic process. HeLa cells were transfected with mtGFP or mtAEQ and then incubated with the indicated doses of TAT wt α-syn. Western blotting (A) and immunolocalization (B) of TAT wt α-syn are shown. Numbers in panel A refer to normalized α-syn/β-actin ratio ± S.E. in three independent Western blottings. Shown is statistical analysis (C) and representative experiments (D) of mitochondrial Ca²⁺ measurements in HeLa cells treated with TAT wt α-syn. Where indicated the cells were stimulated with 100 μm histamine. Bars represent the mean [Ca²⁺] values upon stimulation. Results are the mean ± S.E. ***, p < 0.0005; **, p < 0.005; NS, not significant. The traces are representative of at least four independent experiments. E, shown is Western blot analysis of LC3 I, LC3 II, and p62 levels in TAT α- syn-treated cells; β-actin levels were also shown. Bars indicate the average values obtained by densitometric analysis of four independent experiments. Results are the mean ± S.E. **, p < 0.001.