SNX17 regulates the retromer dependent recycling of Jag1a. (A) Subcellular distribution of SNX17. It was detected at early endosomes (Rab5), late endosomes (Rab7), the retromer complex (Vps35) but not the recycling endosomes (Rab11). (B) The retromer pathway was required for the SNX17-stimulated recycling of Jag1a. Whole cell lysate or the isolated plasma membrane fraction was subject to western blot analysis using the indicated antibodies. Knockdown of Vps35 (35–1, 35–2) but not Rab11 (11–1, 11–2) blocked the SNX17-induced accumulation of Jag1a on the plasma membrane. On the other hand, the recycling of TFR was Rab11 dependent. (C) Knockdown of Vps35 but not Rab11 reduced the Notch reporter activity in 293 T cells. (D) Vps35 was required in the ligand-expressing cells for Notch signaling as determined using the cell co-culture system. Assays are performed as described in Figure 2D. Data represent mean ± SD from three independent assays.