Figure 4.

TAT-mediated delivery of A53T and A30P α-synuclein mutants induces dose-dependent α-syn redistribution and different effect on mitochondrial Ca²⁺ transients. HeLa cells were transfected with mtGFP or mtAEQ and then incubated with the indicated doses of TAT A53T (A) or TAT A30P (B) or TAT WT (C) α-syn. Immunolocalization of TAT mutant α-syn (top) and mitochondrial network (bottom) are revealed by α-syn primary antibodies and mtGFP fluorescence, respectively. The images revealing mitochondrial morphology were randomly acquired from TAT α-syn treated cells. Mitochondrial Ca²⁺ measurements were performed in HeLa cells upon treatment with TAT A53T or A30P or WT α-syn at the indicated doses. Panels on the right show the scatter plots representing the mean [Ca²⁺]_mt peak values upon cell stimulation with histamine. Results are the mean ± SD obtained from at least three independent experiments. *, p < 0.01; **, p < 0.005; ***, p < 0.0005. One-way ANOVA test retrieved a p-value of 0.0001, for multiparametric analysis a Dunnet’s post-test was applied.