Fig. 4.

Dynamin B localizes to the outer mitochondrial membrane. a Dynamin B preprotein localizes to the outside of the outer mitochondrial membrane. Intact mitochondria from AX2 cells overproducing dynamin B-preprotein carrying a C-terminal YFP-tag were purified, treated with trypsin on ice, separated on 8% SDS-PAGE, blotted, and probed with affinity purified rabbit anti-dynamin B antibody or rabbit anti-mNDP kinase. The intermembrane space protein mNDP kinase is not affected, excluding the presence of damaged organelles. The arrows point to the bands for YFP-tagged dynamin B-preprotein, endogenous dynamin B, and mNDP kinase, respectively. b Depletion of dynamin B does not affect mitochondrial morphology. AX2 and dymB⁻ cells were fixed and probed with anti-mitoporin antibody as described above. The superimposition of 30 confocal sections covering the entire cell volume is shown. Scale bars 10 μm