Fig. 5: MAM-Calflux can monitor altered MAM structure and MAM Ca²⁺ levels in Alzheimer’s disease model neurons.

a–d MAM-Calflux detected the decreased MAM Ca²⁺ levels and MAM integrity in the soma of primary cultured 5xFAD tg mouse neurons. Representative images (a) and bar graphs representing BRET ratio (b), MAM area (c), and total Venus excitation intensities (d). (n = 70 for wild-type neurons and n = 76 for 5xFAD tg neurons). e Representative images (left) and Manders’ overlap coefficients (right) of colocalization between mitochondria (TOM20) and ER (Calreticulin) among 5xFAD tg neurons with VAPB-PTPIP51 tethering complex over-expression. (n = 50 for wild-type neurons, n = 43 for 5xFAD tg neurons, and n = 48 for VAPB-PTPIP51 expressing 5xFAD tg neurons). f–i Representative images (f) and graphs representing BRET ratio (g), MAM area (h), and total Venus excitation intensities (i). (n = 48 for wild-type neurons, n = 54 for 5xFAD tg neurons, and n = 54 for VAPB-PTPIP51 expressing 5xFAD tg neurons). Dashed lines in a and f represent neuronal morphologies. The scale bars represent 10 μm. All results are presented as box plots representing the median and ianterquartile 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 h–i are provided as a Source Data file.