Figure 2
From: Mitochondrial proteolytic stress induced by loss of mortalin function is rescued by Parkin and PINK1
Intramitochondrial proteolytic stress phenocopies the reduced viability seen in loss of mortalin function models. (a) Intramitochondrial proteolytic stress in SH-SY5Y cells (n=73) was genetically induced by overexpression of a truncated form of the matrix protein OTC referred to as dOTC. Immunocytochemical studies in fixed cells reveal both OTC and dOTC protein (GFP-green) located within mitochondrial structures shown as Tom20 signal (far red). siRNA transfection was confirmed by fluorescence of the Alexa555 siRNA tag (red). The expressed OTC protein is equally distributed in control cells, but is punctate in mortalin siRNA-treated cells. Quality control experiments to ensure equal expression of OTC and dOTC were performed (data not shown). Scale bars indicate 10 μm. (b) Subsequent ImageJ analysis was done from immunocytochemical studies in Figure 1a. The total area of mitochondria (Tom20 signal) colocalizing with OTC was less in the mortalin knockdown condition compared with the control condition
