Skip to main content
. 2017 Sep 20;10:296. doi: 10.3389/fnmol.2017.00296

FIGURE 4.

FIGURE 4

Endocytosed M6a is sorted to Rab5-positive early endosomes. M6aRFP- expressing neurons at 15 DIV and HEK293 cells were subjected to the antibody internalization assay (AIA). Representative confocal images from M6aRFP-overexpressing cells (shown in green) subjected to the AIA and labeled with anti-Rab5 (shown in magenta). Both neurons (A) and HEK293 cells (B) showed punctate colocalization between M6a and the early endosomal marker, Rab 5, at T30, as shown in the inset in white. Coexpressed M6aRFP/Rab5-GFP neurons (C) and HEK293 cells (D) were subjected to the AIA. Punctate colocalization between both expressed proteins can be seen at T0 and T30 of both types of cells (white in the inset). T0 and T30 represent the steady state and 30 min after immunouptake, respectively. The percentage of colocalization between endocytosed M6a with Rab5 and Rab5-GFP in HEK 293 cells was plotted in (B,D) respectively (for statistical data see Supplementary Table S1). Scale bar: 15 μm. Inset for neurons: 5 × 5 μm; inset for HEK293: 4 × 4 μm.