Figure 6.

Association of SNX1 with Rab6IP1 Is Required for Retromer-Mediated Sorting

(A) GST pull-down of bacterially expressed SNX1 demonstrates that GST-SNX1 can associate with FLAG-tagged Rab6IP1. Comparative levels of GST-SNX1 and GST expression are shown.

(B) Knock down of Rab6IP1 using three different siRNAs and probing for Flag-Rab6IP1 lentivirally transduced into HeLa cells.

(C) Quantification of Rab6IP1 knock down obtained in three different experiments using three different siRNAs. Error bars indicate standard deviation.

(D) Knock down of Rab6IP1 influences the steady-state distribution of CI-MPR, leading to a peripheral dispersal of staining in comparison to control cells, phenocopying loss of retromer components. Knock down of Rab6IP1 does not appear to affect the organization of the TGN (Figure S4).

(E) Quantification of the number of Golgis with more than half-maximal intensity of CI-MPR staining shows that knock down of Rab6IP1 induces a defect in CI-MPR recycling to the TGN. Error bars indicate standard deviation.

(F) Kinetics of the trafficking of CD8-CI-MPR from the cell surface to the TGN shows that loss of Rab6IP1 reduces the efficiency of retromer-mediated transport in comparison to control cells. Error bars indicate standard deviation.