Figure 2

UGT1A1 binds to UGT1A1 or UGT1A1-p.Q331X. (a) UGT1A1 binds to UGT1A1 in vitro. Lane 1: His-UGT1A1 and MBP-UGT1A1 expressed in E. coli strain BL21 (DE3). Lane 2: SDS eluate of His-bound Ni²⁺ beads following incubation with MBP-UBT1A1. Lane 3: SDS eluate of His-UGT1A1-bound Ni²⁺ beads following incubation with MBP-UGT1A1. (b) UGT1A1 dimerization analysis using AFM. In some of the particles the right-hand side in the dimer was larger than the left-hand side. This configuration may have been caused when the buffer was evaporated by helium gas. (c) UGT1A1 binds to UGT1A1-p.Q331X in vitro. Lane 1: His-UGT1A1-p.Q331X and MBP-UGT1A1 expressed in E. coli strain BL21 (DE3), Lane 2: SDS eluate of His-bound Ni²⁺ beads following incubation with MBP-UGT1A1, Lane 3: SDS eluate of His-UGT1A1-p.Q331X-bound Ni²⁺ beads following incubation with MBP-UGT1A1. The Ni²⁺ pull-down assay was performed using His-UGT1A1 or His-UGT1A1-p.Q331X as the bait. Eluates were analyzed by SDS-PAGE followed by western blotting using a polyclonal anti-UGT1A1 antibody. A full color version of this figure is available at the Journal of Human Genetics journal online.