EGFP-Rab5 (Addgene) was a gift from Arnab Gupta. Cells were transfected with 1 μg of Rab5 plasmid DNA to label early endosomes, respectively, by using Lipofectamine 3000 (Invitrogen). EGFP-CAAX [41 (link)] was a gift from Lei Lu. Cells were transfected with 1 μg EGFP-CAAX (Addgene) plasmid to mark the membrane. Cells were transfected with 1 μg of mCherry- Clathrin LC-15 (Addgene) to label clathrin-coated vesicles. To mark fast recycling endosomes, cells were transfected with 1 μg of mCherry- Rab 4a plasmid. Other treatments, if required, were performed 16 h after transfection.
Fixation of cells was performed by using 4% paraformaldehyde (Sigma-Aldrich) for 15 min at 37°C temperature.
Fixation of cells was performed by using 4% paraformaldehyde (Sigma-Aldrich) for 15 min at 37°C temperature.
Free full text: Click here
