Fig. 6.

MAM disruption in mutant FUS iPSC-derived OPCs. a Immunofluorescent staining for DAPI, VAPB, and PTPIP51 on DIV26-30 OPCs generated from FUS^P525L, FUS^R521H and their isogenic control hiPSCs (scale bar: 10 μm). b, d Violin plot of the intensity of VAPB staining on FUS^P525L and FUS^R521H OPCs, and their isogenic controls. Each symbol represents the mean intensity of VAPB from an image. Data are represented as mean ± SD from 3 biological (N = 30 images). c, e Violin plot of intensity of PTPIP51 staining on FUS^P525L and FUS^R521H OPCs, and their isogenic controls. Each symbol represents the mean intensity of PTPIP51 from an image. Data are represented as mean ± SD from three biological (N = 30 images). f Proximity ligation analysis (PLA) using antibodies against VAPB and PTPIP51 (VAPB/PTPIP51, red) FUS^P525L and FUS^R521H OPCs, and their isogenic controls. Nuclei were stained with DAPI (blue). Scale bars are 10 μm. g, h Violin plot of quantification of the PLA signal in FUS^R521H and isogenic control OPCs (h) and FUS^P525L and isogenic control OPCs (i). Each symbol represents the mean number of PLA dots per cell from one image. Statistical analyses were performed by unpaired two-tailed t tests to compare mutant FUS OPCs and their controls. Data are represented as mean ± SD from three biological replicates with three technical replicates in each experiment (N = 20 images and n > 100 nuclei/condition). **p < 0.01