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Image Search Results
Journal: EMBO Reports
Article Title: APEX2‐mediated RAB proximity labeling identifies a role for RAB21 in clathrin‐independent cargo sorting
doi: 10.15252/embr.201847192
Figure Lengend Snippet: RAB21 is required for endosomal sorting of direct clathrin‐independent cargos. RAB21 associates with WASH/retromer subcomplexes at endosomes. RAB21 could either (i) recruit WASH/retromer or (ii) be recruited by WASH/retromer or (iii) be part of a positive feedback loop that would allow WASH/retromer and RAB21 recruitment at endosomes. Endosomal RAB21 would be required for WASH‐mediated F‐actin polymerization. Although the data do not directly demonstrate a direct link between F‐actin generation and cargo sorting, we propose that RAB21‐dependent F‐actin generation would be required for sorting of a CIE cargo subclass (MCT1, SLC3A2, Basigin, and CD44), while it would not be required for other cargos (CI‐MPR or Glut1). In RAB21 knockout cells, decreased WASH/retromer endosomal localization is observed, which results in reduced endosomal F‐actin and direct CIE cargo misrouting. This ultimately leads to the lysosomal degradation of these misrouted cargos (demonstrated for SLC3A2). Dotted line between VARP and RAB21 indicates that their coregulation with the retromer is speculative.
Article Snippet: Antibodies used for immunoblotting were anti‐GFP (1:500, Roche #11814460001), anti‐RAB21 (1:1,000, Sigma #R4405 or 1:1,000, Invitrogen #PA5‐34404), anti‐RAB4 (1:1,000, Cell Signaling #2167), anti‐RAB5 (1:1,000, Cell Signaling #3547), anti‐RAB7 (1:1,000, Cell Signaling #9367), anti‐GAPDH‐HRP (1:1,000, Cell Signaling #8884), anti‐Myc (1:1,000, Cell Signaling #2278), anti‐HA (1:1,000, Cell Signaling #3724),
Techniques: Knock-Out