Research Article
Disruption of Intracellular Calcium Homeostasis Leads to ERLIN2-Linked Hereditary Spastic Paraplegia in Patient-Derived Stem Cell Models
Figure 4
The ERLIN2 p.V71A mutant increased the apoptosis of NSCs via ER stress and can be rescued by TUDCA. (a) Apoptosis was detected by flow cytometry after staining with Annexin V-FITC apoptosis detection kit. The early apoptosis percentage was expressed as Q3. The bar chart represents the quantification of the result (, experiments, ) with iPS-GDL as the WT and iPS-GJH as the p.Val71Ala. (b) Western blot showed that caspase 3, cleaved-caspase 3, caspase 12, and CHOP were significantly upregulated (, experiments, , ). (c) The representative EM images and quantification of mitochondria number and the ER lumen/area in WT and p.Val71Ala NSCs (, images, samples, , ), iPS-GDL as the WT group and iPS-GJH as the p.Val71Ala group. The red arrows represent the ER, and the yellow arrows represent the mitochondria. (d) The iPS-GJH-derived NSCs were treated with 100 μM DMSO and 100 μM tauroursodeoxycholate (TUDCA). Apoptosis was detected by flow cytometry after staining with Annexin V-FITC apoptosis detection kit. The early apoptosis percentage was expressed as Q3. The bar chart represents the quantification of the result (, experiments, ). (e) Western blot showed that the protein level of caspase 3 was not significantly different, while the cleaved-caspase 3 and caspase 12 had been significantly downregulated when the cells were treated with TUDCA (, experiments, , , ).
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