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. 2021 Jun 25;297(2):100921. doi: 10.1016/j.jbc.2021.100921

Figure 3.

Figure 3

Mapping the region of TDP1 that interacts with LigIII.A, pull-down assay with N-terminal truncated (Δ148) single amino acid mutant TDP1. Upper panel, input (20%) of the indicated proteins. Immunoblotting with (left gel) rabbit anti-TDP1-C-term (Abcam) and (right gel) anti-GST (Santa Cruz Biotechnology). Lower panel (left gel), Ni-NTA beads liganded by LigIIIβ (10 pmol) were used to pull down the indicated proteins (10 pmol of each). Reaction controls (right gel) were Ni-NTA beads liganded by LigIIIβ incubated with GST and Ni-NTA beads alone incubated with GST-TDP1. Immunoblotting with rabbit anti-TDP1-C-term (Abcam). B, summary of pull-down assays with N- and C- terminally-truncated versions of TDP1. Asterisks represent the relative efficiency of the interaction.