Xenbase Image ID: 156419

Figure 6. XPF leucine 219 is part of the major interaction site between XPF and SLX4 pICL was replicated in XPF‐ERCC1‐depleted (ΔXE) extract or in XPF‐ERCC1‐depleted extract supplemented with wild‐type XPF‐ERCC1 only (+XEWT), wild‐type XPF‐ERCC1 and SLX4 (+SXEWT), or XPFL219R‐ERCC1 and SLX4 (+SXEL219R; see Fig EV5A). Samples were taken at the indicated times and immunoprecipitated with α‐XPF (left panel) or α‐SLX4 antibodies (right panel). Co‐precipitated DNA was isolated and analyzed by quantitative PCR using ICL or pQuant primers. The qPCR data were plotted as the percentage of peak value with the highest value set to 100%. Wild‐type and mutant FLAG‐XPF‐ERCC1 were co‐expressed with His‐SLX4 in Sf9 insect cells. Cells were lysed and XPF was immunoprecipitated via the FLAG‐tag. Samples were analyzed by Western blot using α‐FLAG and α‐His antibodies. In, input; FT, flow‐through fraction; B, fraction bound to beads. Schematic representation of xlSLX4 proteins, with the MLR and BTB domains indicated. Experimental replicates are shown in Fig EV5. Purified wild‐type FLAG‐SLX4 and FLAG‐SLX4∆MLR were added to Xenopus egg extract. SLX4 was immunoprecipitated via the FLAG‐tag. Samples were analyzed by Western blot using α‐FLAG and α‐XPF antibodies. Line within blot indicates position where irrelevant lanes were removed. *, background band. Source data are available online for this figure. Image published in: Klein Douwel D et al. (2017) © 2017 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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