Fig. 1
A loss of XPB NTD integrity induces large-scale chromatine decondensation. a Schematic representation of the lacO/LacR tethering system used in U2OS17 cells. See the Materials and methods section for a full description of the cell line. b Schematic representation of wild-type and mutant XPB-LacR-GFP constructs. For clarity, the sizes of the GFP (238 aa) and LacR (367 aa) are omitted. c Proteins from whole-cell extracts (15 μg) of U2OS17 cells transiently transfected with wild-type or mutant XPB-LacR-GFP constructs were resolved by SDS-PAGE and immunoblotted using either polyclonal rabbit anti-GFP (upper panel), polyclonal rabbit anti-XPB (middle panel) or monoclonal mouse anti-Actin antibodies (lower panel). Source data are provided as a Source Data file. d U2OS17 cells were transiently transfected with 1 μg of expression vectors for the following proteins: LacR-GFP, XPBWT-LacR-GFP, XPB320–782-LacR-GFP, XPBF99S-LacR-GFP, XPBT119P-LacR-GFP, XPB1–550-LacR-GFP. In parallel, U2OS17 cell line was transiently transfected with 5 μg of expression vector for LacR-GFP. GFP was observed by fluorescence microscopy 24 h post transfection. Lower panels are magnifications of the white rectangles in the upper panels. e 3D reconstruction of U2OS17 cellular nuclei transiently transfected either with XPBWT-LacR-GFP or with XPBF99S-LacR-GFP using Imaris Software (Bitplane). f Whisker box plot shows quantifications of the relative array volumes (volume of the array/volume of the nucleus; 40–150 cells for each condition). Significant p-values are indicated (*** ≤ 0.001) and were obtained using a Kruskal Wallis test. Source data are provided as a Source Data file
