Fig. 3: MAM-Calflux measures changes in the steady-state Ca2+ level at MAM.
From: Ratiometric measurement of MAM Ca2+ dynamics using a modified CalfluxVTN
a–d Increased MAM Ca2+ levels and enhanced MAM structure formation measured by MAM-Calflux upon over-expression of VAPB-PTPIP51 tethering complex in HeLa cells. Representative images (a) and bar graphs representing BRET ratio (b), MAM area (c), and total Venus excitation intensities (d). Dashed lines in a represent the cell morphologies. (n = 45 for control HeLa cells, n = 33 for VAPB-PTPIP51 expressing HeLa cells). e–h Increased MAM Ca2+ levels and intact MAM structure measured by MAM-Calflux upon over-expression of GRP75 in HeLa cells. Representative images (e) and bar graphs representing BRET ratio (f), MAM area (g), and total Venus excitation intensities (h). (n = 68 for control HeLa cells, n = 82 for GRP75 expressing HeLa cells). i–l Decreased MAM Ca2+ levels and reduced MAM structure formation measured by MAM-Calflux upon knockdown of MFN2 in HeLa cells. Representative images (i) and bar graphs representing BRET ratio (j), MAM area (k), and total Venus excitation intensities (l). (n = 108 for control HeLa cells, n = 99 for MFN2 knockdown HeLa cells). Dashed lines in a, e, and i outline the cell 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, f–h, and j–l. Source data from b–d, f–h, and j–l are provided as a Source Data file.
