Figure 6. Pre-treatment of EPO maintains intracellular calcium and prevents MPTP opening in H/R induced H9C2 cells.

The effect of EPO on intracellular calcium and MPTP opening was determined using time–lapse imaging in confocal microscope. (A) H9C2 cells were subjected to 8 hrs hypoxia and 15 mins reperfusion with or without pre-treatment with 20 U/ml EPO for 24 hrs and stained with Fluo-4 AM and performed live cell imaging. H9C2 cells pretreated with EPO accumulate Ca²⁺ inside the mitochondrial membrane even after reperfusion and showed increase Fluo-4 AM fluorescence. H9C2 cells subjected to H/R showed release of Ca²⁺ outside the mitochondria due to overload of Ca²⁺ during reperfusion rupture of mitochondrial membrane occurs. (B) Represents the quantification of Fluo-4 AM fluorescence. Data are presented as means ± SEM of the ratios from three independent experiments * denotes p<0.001 for analyses compared to H/R.