Fig. 6: MAM-Calflux can determine the altered MAM-calcium physiology in the α-syn-mediated Parkinson’s disease model neurons.

a–d MAM-Calflux detected the increased MAM steady-state Ca²⁺ levels with diminished structure integrity in primary cultured SNCA*A53T tg mouse neurons at DIV7. Representative images (a) and bar graphs representing BRET ratio (b), MAM area (c), and total Venus excitation intensities (d). (n = 71 for wild-type neurons and n = 66 for SNCA*A53T tg neurons). e Representative images (left) and Manders’ overlap coefficients (right) of colocalization between mitochondria (TOM20) and ER (Calreticulin) among SNCA*A53T tg neurons with VAPB-PTPIP51 tethering complex over-expression. (n = 55 for wild-type neurons, n = 43 for SNCA*A53T tg neurons, and n = 36 for VAPB-PTPIP51 expressing SNCA*A53T tg neurons). f–i Representative images (f) and graphs representing BRET ratio (g), MAM area (h), and total Venus excitation intensities (i). Dashed lines in f represent the neuronal morphologies. (n = 77 for wild-type neurons, n = 95 for SNCA*A53T tg neurons, and n = 92 for VAPB-PTPIP51 expressing SNCA*A53T tg neurons). Dashed lines in a and f represent the neuronal morphologies. The scale bars represent 10 μm. All results are presented as box plots representing the median and interquartile range with whiskers min/max value and the cross representing the mean value. All P values were calculated using two-tailed Student’s t test for b–d and one-way ANOVA with Bonferroni’s multiple comparison tests for e and g–i. Source data from b–e and g–i are provided as a Source Data file.