Extended Data Fig. 7: Migrasomes transport GPR56 across cells.
From: Adhesion GPCR-induced ectocytosis mediates intercellular GPCR signal propagation
a, Left panel: Schematic diagram illustrating the pH-dependent fluorescence of the pHluorin tag. The C-terminally tagged pHluorin on GPR56 fluoresces at the plasma membrane and in early endosomes, where the pH is neutral, but is quenched in acidic compartments such as lysosomes and multivesicular body. Right panel: Representative confocal images of HEK293-GPR56-pHluorin cells co-cultured with HEK293T cells expressing the membrane marker CAAX-mCherry for 12 hours. Scale bar, 10 μm. Insets show enlarged regions of interest (ROIs). b, HEK293T cells were incubated for 24 hours with purified migrasomes derived from HEK293GPR56-pHluorin cells, along with the treatment of DMSO (control), 50 μM dynasore, or 30 μM Pitstop2. Cells were stained with Hoechst and then imaged by confocal microscopy. Scale bar, 10 μm. Quantification of uptake of GPR56-positive puncta per cell is shown in the lower left panel. Data are presented as mean ± SEM from over 30 cells in three biologically independent experiments. Statistical significance was determined by ordinary one-way ANOVA with Tukey’s test. ****P < 0.0001
