Journal: Biochemistry and biophysics reports

Article Title: Acetylation of the nuclear localization signal in Ku70 diminishes the interaction with importin-α.

doi: 10.1016/j.bbrep.2022.101418

Figure Lengend Snippet: Fig. 1. Effect of substitution of lysine residues in the Ku70 NLS with ace tyl–lysine on binding to Impα. (A) The effect of amino acid substitutions in the Ku70 NLS on Impα binding. The binding activity of Ku70 NLS and its mutant peptides to Impα in the presence of Impβ was analyzed by probing immunoblots with antibodies against Impα. (B) Quantification of the pull-down assays pre sented in panel (A). Measurements of the blot band intensity were performed using ImageJ 1.52a. Each graph represents the relative intensity with Ku70 NLS WT defined as 100%. The error bars indicate the standard deviation from three independent experiments. *p < 0.05 significant differences from the case when the expected value was defined as 100.

Article Snippet: A binding assay was performed with 50 μl NLS-immobilized sepharose, 1 mg/ml bovine serum albumin, 0.1 μg recombinant human Impα2 (NBP1-78888; Novus Biologicals, Centennial, CO, USA), and 0.1 μg recombinant human Impβ (NBP1-78815; Novus Biologicals) in 0.5 ml transport buffer by incubation for 2 h at 4 ◦C with gentle rotation.

Techniques: Binding Assay, Activity Assay, Mutagenesis, Western Blot, Standard Deviation