Figure 5.

PLAUR/TLR4 interaction increases DNA repair efficiency. (a) PLAUR was immunoprecipitated from HeLa cell lysates after Dox treatment, TLR4 was detected in the immunoprecipitates by western blotting. Normalization was performed by the amount of TLR4 present in the original lysates. The experiment was repeated in the reverse direction (Right). (b) HeLa cells were treated with radiation of 9 Gy and fixed at the indicated time points, Duolink proximity ligation assay was performed to determine the interaction between PLAUR and TLR4. Duolink signal per cell was counted and represented as a graph (right). Scale bar 20 μm. Data shown as mean ±S.D. (c) DNA repair assay using plasmid substrates pDRGFP (for HR) and pEJ2GFP (for NHEJ), in WT and PLAUR-expressing HEK-Blue hTLR4cells having constitutive expression of TLR4, CD14 and MD-2. Cells were transfected with pCBASceI for the introduction of a double-strand break and DNA repair efficiency correlating with GFP expression was assessed after 3 days by FACS. Data shown as mean ±S.D.