Skip to main content
. 2012 Jul 16;109(31):12485–12490. doi: 10.1073/pnas.1207821109

Fig. 4.

Fig. 4.

DRD3 is involved in endocytic sorting. (A) DRD3 RNAi causes increased CI-M6PR localization to vesicles in addition to the Golgi. Control or DRD3 RNAi-treated cells were stained for CI-M6PR (green) and a resident Golgi marker GM130 (red). (B) Pulse and chase experiments show sorting defects in DRD3 RNAi cells. HeLa cells were treated with control or DRD3 RNAi for 3 d before Alexa 555-EGF, FITC-70 kD-dextran, and Alexa 647-Transferrin were added for 10 min followed by washing out with marker- and phenol red-free medium. Pictures were taken at indicated time points in live cells. (C) Quantification of EGF and/or dextran that colocalize or partially colocalize with transferrin; (D) Endocytic cargoes are trapped in SNX1-positive endosomes after DRD3 RNAi. HeLa cells were treated with control or DRD3 RNAi for 3 d before Alexa 555-EGF (red) was added for 10 min followed by washing out with marker-free medium for indicated time points. Cells were than washed, fixed, and stained with anti-SNX1 (green). EGF that colocalizes or partially colocalizes with SNX1 was quantified. For each condition, around 50 cells were counted. Mean values from two independent experiments are shown. (Scale bar: 10 μm.) (E) Flow cytometry analysis of transferrin recycling in control or DRD3 RNAi knockdown HeLa cells.