Fig. 5
Rapid Screening of a GFP-Tagged Phosphoinositide Probe Library using APEX-GBP Allows for Fast and Reliable Protein Localization
(A?H) GFP-PLC-PH, GFP-BTK-PH, GFP-AKT-PH, GFP-TAPP1-PH, GFP-2xFYVEhrs, GFP-OSBP-PH, GFP-2ML1N and GFP-2ML1N (7Q) detected at known subcellular localizations by APEX-GBP co-transfection. Scale bars represent 1 µm. Untransfected adjacent cells without reaction product. Arrowheads represent enriched electron density at the PM, arrows highlight accumulation of reaction product within endosomes. Cy, cytoplasm; PM, plasma membrane; EEs, early endosomes; LE, late endosomes; Golgi, Golgi complex; M, mitochondria; CCP, clathrin-coated pit. The above localizations were confirmed by confocal microscopy; see Figure S3. Comparisons of ultrastructure between modular and directly APEX-tagged PI probes were performed; see Figures S4A?S4C.
Reprinted from Developmental Cell, 35(4), Ariotti, N., Hall, T.E., Rae, J., Ferguson, C., McMahon, K.A., Martel, N., Webb, R.E., Webb, R.I., Teasdale, R.D., Parton, R.G., Modular Detection of GFP-Labeled Proteins for Rapid Screening by Electron Microscopy in Cells and Organisms, 513-25, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell