|
|
|
|
|
||
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 8. BCS1L downregulation causes supercomplex disassembly and abnormal mitochondrial morphology. (A) Total cell lysates from cells transfected with siRNA targeting GFP (control) or BCS1L were immunoblotted with antibodies to the indicated proteins. BCS1L knockdown caused reduction in the supercomplexes of the respiratory chains. (B) Mitochondrial fractions from cells transfected with siRNA targeting GFP (control) or BCS1L were solubilized with digitonin, subjected to Blue-Native gel electrophoresis, and immunoblotted using antibodies against the indicated proteins. Arrowhead indicates supercomplexes of Complex I, III, and IV. Asterisk indicates a nonspecific band. (C) HeLa cells that stably expressed Su9-DsRed were transfected with siRNA for GFP (panel a), BCS1L (panels b,c,d), LETM1 (panel e), or BCS1L and LETM1 (panel f), and further transfected with plasmid co-expressing BCS1L and nucleus-targeted GFP (panel b). BCS1L knockdown induced abnormal mitochondrial morphology. Live images were acquired by confocal microscopy. Typical images are shown in insets at high magnification. Scale bar, 10 µm. (D) Cells transfected with BCS1L siRNA were analyzed by immunofluorescence microscopy. Data represent the mean ± s.e. of three independent experiments; 100-200 individual cells were counted. (E) Model of the functions of BCS1L and LETM1 in mitochondrial biogenesis. Blue arrows indicate functions reported here; gray arrow indicates the BCS1L function reported previously (Cruciat et al., 1999; de Lonlay et al., 2001).
|
