Figure 2.

Renal I/R stress increases RIP3 interaction with Mitofilin in mitochondria. (A) Immunoblots showing the link between RIP3 and Mitofilin in the inner mitochondrial membrane of HK-2 cells. Immunoprecipitation (IP) in whole-cell lysate fractions with myc-Mitofilin antibody immunoprecipitated flag-RIP3, which was revealed by Western blot analysis. Reverse Co-IP with flag-RIP3 confirmed the interaction between these two proteins, as myc-Mitofilin was detected in the immunoprecipitate; n = 3 independent experiments. (B) Confocal microscopy images of kidneys from sham animals and after AKI labeled with RIP3 (green) and Mitofilin (red) and the overlay of both proteins (yellow). The graph shows the increase in RIP3 and the reduction in Mitofilin levels after AKI compared with sham kidneys. These results indicate that the co-localization between Mitofilin and RIP3 is increased after AKI (60 min ischemia followed by 12 h reperfusion); * p < 0.05 versus sham group, respectively, n = 3 experiments.