Figure 4

Location and function of TMF A. Immunoblot of Golgi and cytosol fractions with anti-TMF antibodies. The Golgi lane contained approximately 5 μg total protein, the cytosol lane approximately 100 μg protein. Marker positions (kDa) are indicated. B. Immunofluorescent images of COS cells showing endogenous TMF in an untransfected cell (untransf.), HA-tagged full-length protein expressed from a transfected plasmid (HA-TMF) and detected with both anti-HA and anti-TMF, as well as a cell expressing rather higher levels of HA-TMF (only the anti-HA image is shown; anti-TMF staining of this cell gave an identical pattern). The last panel shows the HA-tagged 310 C-terminal residues of HA (HA-C term) expressed similarly and detected with anti-HA. Bar is 10 μm. C. Rat NRK cells transfected with RNAi and stained for TMF and either TGN38 or GOS28 as indicated. Note that TMF-depleted cells (green in the merged images) have more dispersed Golgi than the other cells. The insets in some panels show an enlarged region of the Golgi, boxed in the merged image. Bars are 10 μm.