Electron microscopy of inclusion bodies in 293 cells expressing mycRab24(D123I). Cells were harvested 24 h after transfection. Expressed protein was detected with the mouse monoclonal antibody against the myc epitope, followed by secondary antibody conjugated to 10 nm gold particles. Cells were processed for electron microscopy as described in the Methods. Panel A shows a cell where the majority of immunoreactive inclusions accumulated within nucleus (N). Panel B shows the intranuclear inclusions at higher magnification, revealing the individual gold particles concentrated in dense spherical particles of nearly uniform diameter. Panel C shows an example of a cell where disruption of the nuclear envelope was observed (between arrows) in the region near the accumulated intranuclear inclusions. Panel D shows a cell where inclusions were observed in the perinuclear cytoplasm. The dimension bars in each panel are as follows: A, 0.5 micron; B, 0.1 micron; C, 0.5 micron; D, 0.5 micron. The solid arrowheads in panels A, C and D indicate the nuclear membrane.
Maltese et al. BMC Cell Biology 2002 3:25 doi:10.1186/1471-2121-3-25