|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 5. Polarized trafficking of biosynthetic cargo to pseudopods in prostate tumor cells. R3327-5'A cells were transfected with plasmids encoding GFP-VSVG (tsG-GFP) or GFP 
5-integrin (5-GFP) and morphological transport assays were performed as described in the Materials and Methods. Following accumulation of cargo proteins in the TGN, cultures were shifted to 32°C either in the absence (A) or presence (B) of 0.5% tannic acid for various lengths of time to facilitate the trafficking of accumulated GFP-fusion proteins from the TGN to the plasma membrane. After 30 minutes at 32°C (A) or indicated times (B), cells were fixed with 2% paraformaldehyde, permeabilized and stained with anti-Sec8 mAb, which was detected using a Texas Red-conjugated secondary antibody. Epifluorescence images were obtained as described in Fig. 1. Arrowheads indicate TGN and arrows indicate examples of post-TGN transport intermediates. More post-TGN transport vesicles appear to be delivered to pseudopods than to other parts of the cell, even when the TGN is located on the opposite side of the nucleus from the pseudopod. In tannic acid pre-fixed samples (B), pseudopods accumulate transport vesicles, but with prolonged incubation, vesicles also begin to accumulate within the cell body.
|
