T6BP and NDP52 colocalise with myosin VI in vesicles in the perinuclear region around the Golgi complex in HeLa cells. (A) Cells transiently overexpressing GFP-tagged myosin VI tail (a-d, e-h) were pre-permeabilised with saponin, fixed, double labelled with the antibodies against T6BP (a-d) and NDP52 (e-h), and analysed by immunofluorescence. The boxed areas in panels a and e are shown enlarged in panels panels b-d and f-h, respectively. Arrows indicate examples of colocalisation. Bars, 10 μm. (B) Immuno-electron microscopy shows endogenous T6BP (a,b) and endogenous NDP52 (c,d). Myosin VI and T6BP (b) or NDP52 (d) colocalise in vesicles near the Golgi complex. The number of gold particles observed in these sections reflects the expected levels of these endogenous proteins. Enlarged images highlight T6BP and NDP52 associated with vesicles. Bar, 450 nm.