Fig. 5
Increased sensitivity to opening of the mPTP in OPA1 iPSC-RGCs in response to mitochondrial calcium overload. (a) Representative live cell confocal images of the MMP in Mitotracker-stained isogenic control and R445H iPSC-RGCs at baseline and after treatment with 12.5 µM and 20 µM ferutinin. Scale bar, 20 μm. (b & c) Quantification of fold-change in Mitotracker fluorescence over time in response to mitochondrial calcium overload in isogenic control and R445H neurons. Fluorescence intensity was normalised to baseline values (F0). Ferutinin concentration was increased in 2.5 µM increments at the indicated time points indicated by the arrows. Loss of Mitotracker fluorescence indicates mitochondrial depolarisation due to opening of the mPTP (grey box). (d) Quantification of ferutinin concentration required to open the mPTP. Isogenic control/R445H, n = 7 experiments (~ 350 cells/genotype) sampled from four independent differentiations. ** = p < 0.01, unpaired t-test. Scatter plot shows the mean values +/- SD