Fig. 1.

Pull-down of human sodium-dependent multivitamin transporter (hSMVT) from Caco-2 cells by GST-tagged PDZD11. A: generation and purification of GST-PDZD11. GST-PDZD11 fusion protein was produced in BL-21 Escherichia coli cells and purified as described in experimental procedures. Protein samples were run on NuPAGE 4–12% Bis-Tris gradient gels and stained with Coomassie blue. B: pull-down of hSMVT from Caco-2 cells by GST-PDZD11 but not by GST alone. Postnuclear extract of Caco-2 cells (1 mg of total soluble protein) was incubated with either GST-PDZD11 (100 μg) or GST (100 μg) bound to glutathione-Sepharose4B beads. Bound proteins were eluted with 10 mM glutathione and analyzed by Western blotting using the anti-hSMVT antibodies.