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Fig. 1. Mitochondria are partially localised along microtubules in live HeLa cells. (A) Cells were co-transfected with 0.25 µg mito.DsRed and 1 µg -tubulin.EGFP. Forty-eight hours after transfection cells were imaged on an UltraVIEW^TM Live Cell Confocal Imaging system in KRH buffer (see Materials and Methods) at 22°C. The boxed region in a is shown on an expanded scale in b); arrows in b indicate regions of colocalisation. (B) Microtubules were depolymerised with 10 µM nocodazole. The boxed region in a is shown on an expanded scale in b. Bars, 1.0 µm. (C) Dynactin subunits (p50 and p150^Glued) and dynein are associated with a mitochondrial fraction. Heavy mitochondrial pellet (HMP) was obtained after spinning the postnuclear supernatant (PNSN) at 3000 g for 10 minutes (see Materials and Methods). An equal amount (3 µg) of protein was loaded each lane. The intensity of the protein bands was measured using NIH ImageJ software (http://rsb.info.nih.gov/ij/). PHMSN, post-heavy mitochondrial supernatant; DIC, dynein intermediate chain.

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