Skip to main content
. 2010 Apr 19;107(18):8225–8230. doi: 10.1073/pnas.1000157107

Fig. 3.

Fig. 3.

The effect of PtdIns4P elimination on mannose 6-phosphate receptor trafficking in COS-7 cells. Cells were transfected with the CI-M6PR fused to photoactivable-GFP (A Left) either alone or together with the Golgi-targeted Tgn38-FRB (nonfluorescent) and the cytosolic mRFP-FKBP-only (A Middle) or the cytosolic mRFP-FKBP-Sac1 phosphatase (A Right). The next day the mounted live cells were preincubated for 10 min with rapamycin (100 nM) and the Golgi region of cells that showed recruitment was photoactivated. Time-lapse images were followed for 5 min with GFP excitation. (A) Representative images of each group of cells at the indicated time points. (B) Combined result of morphometric analysis of the recordings using MetaMorph software. The graphs show the number of objects around the Golgi that are above an arbitrary threshold normalized to the value found right after photoactivation (at 0 min). The graph illustrates the first 2.5 min of the recordings. (Means ± SEM of 12, 12, and 21 cells for the black, gray, and open symbol traces, respectively.)