Fig. 3.

Pull-down assay of hRFC from human intestinal epithelial Caco-2 cells by GST-DYNLRB1. A: generation and purification of GST-DYNLRB1 fusion protein and GST. Extracts from BL-21 Escherichia coli cells harboring recombinant pGEX-4T-1 (1), purified GST (2), pGEX-4T-1-DYNLRB1 (3), purified GST-DYNLRB1 (4), and marker (M). Protein samples were run on SDS-PAGE and stained with Coomassie blue. B: Caco-2 cells lysate was incubated with glutathione-Sepharose 4B beads adsorbed with GST-DYNLRB1 or GST alone (negative control). Proteins bound to the beads were separated, washed and analyzed by Western Blotting using anti-hRFC antibodies. 1, Caco-2 cell lysate; 2, proteins bound to GST-DYNLRB1; 3, proteins bound to unfused GST.