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. 2020 Jul 7;32(1):107850. doi: 10.1016/j.celrep.2020.107850

Figure 4.

Figure 4

Ubiquitination of WRNIP1 Is Required for Recruitment to ICLs

(A) His-tagged WRNIP1 was purified from Sf9 insect cells and analyzed by immunoblot analysis using the antibodies indicated.

(B) Coomassie blue staining of Flag-HA-WRNIP1 purified together with associated proteins from HeLa S3 cells.

(C) Magnified view of part of (B), showing the area of the gel that was excised for mass spectrometry analysis.

(D) Schematic showing locations of amino acids in WRNIP1 that were found to be modified by ubiquitin.

(E) Schematic showing locations of lysines in WRNIP1 that were mutated into arginines in WRNIP1-19R.

(F) Live-cell imaging of HeLa-WRNIP1-EGFP and HeLa-WRNIP1-19R-EGFP cells. Cells were treated with TMP, microirradiated at the indicated areas (white arrows), and followed over time. Quantification shows relative intensities of the irradiated areas. Data represent 2 independent experiments (n = 2), and 10 cells were quantified for each chart (mean ± SEM). Scale bar, 10 μm.