Figure 7. TPD54 co-reroutes dileucine motif-containing receptors only. (A) Representative widefield micrographs of cells coexpressing mCherry-FKBP-TPD54, dark MitoTrap, and the indicated CD8 construct. Rerouting was induced by 200 nM rapamycin. Cells were fixed, permeabilized, and stained for total CD8. (B) Representative widefield micrographs showing co-rerouting of endogenous CIMPR detected by immunofluorescence with rerouting of mCherry-FKBP-TPD54 to dark MitoTrap by addition of 200 nM rapamycin. (C) Pulse label and timed vesicle capture experiments. Cells expressing CD8-EAAALL were surface labeled with Alexa Fluor 488–conjugated anti-CD8 antibodies for 30 min, then incubated at 37°C for the indicated time (minutes), treated with 200 nM rapamycin for 5 min, and fixed. (D) Representative widefield micrographs from a pulse label and timed vesicle capture experiment. Inset, 5× zoom. Scale bars, 10 µm, 1 µm (insets). Time, hours and minutes.
Figure 7.

TPD54 co-reroutes dileucine motif-containing receptors only. (A) Representative widefield micrographs of cells coexpressing mCherry-FKBP-TPD54, dark MitoTrap, and the indicated CD8 construct. Rerouting was induced by 200 nM rapamycin. Cells were fixed, permeabilized, and stained for total CD8. (B) Representative widefield micrographs showing co-rerouting of endogenous CIMPR detected by immunofluorescence with rerouting of mCherry-FKBP-TPD54 to dark MitoTrap by addition of 200 nM rapamycin. (C) Pulse label and timed vesicle capture experiments. Cells expressing CD8-EAAALL were surface labeled with Alexa Fluor 488–conjugated anti-CD8 antibodies for 30 min, then incubated at 37°C for the indicated time (minutes), treated with 200 nM rapamycin for 5 min, and fixed. (D) Representative widefield micrographs from a pulse label and timed vesicle capture experiment. Inset, 5× zoom. Scale bars, 10 µm, 1 µm (insets). Time, hours and minutes.

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