Enhanced mitochondrial biogenesis promotes neuroprotection in human pluripotent stem cell derived retinal ganglion cells - Communications Biology

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Enhanced mitochondrial biogenesis promotes neuroprotection in human pluripotent stem cell derived retinal ganglion cells - Communications Biology
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. Next, hRGCs were resuspended in iNS media, counted using a hemocytometer and seeded on MG-coated plates, coverslips, or MatTek dishes for experiments.Purified hRGCs were seeded at 30,000 cells per well of a 96-well MG-coated plate and maintained for 3 days. For measuring mitochondrial mass, hRGCs were labeled with mitochondria-specific MTDR dye. To measure mitochondrial degradation, hRGCs were labeled with 10 nM MTDR dye for 1 h, washed, and then treated with 10 μM CCCP for a time course.

. Protein bands corresponding to the appropriate size were quantified following product datasheets and published literature.Purified hRGCs were seeded on MG-coated coverslips at a density of 30,000–40,000 cells per coverslip and grown for 3 days. Next, hRGCs were treated with indicated molecules and time points. After treatment the media was aspirated, the cells were washed with 1× PBS, and then fixed with 4% Paraformaldehyde for 15 min at 37 °C. Fixed cells were permeabilized with 0.

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