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. 2016 Sep 29;7(9):e2383. doi: 10.1038/cddis.2016.291

Figure 4.

Figure 4

PLAUR/TLR4 signaling is important during DNA damage. (a) Normal HeLa cells were treated with 2 μM Dox for 1 h, washed and conditioned media (CM) was collected after 3 h. This CM was added to sicon and PLAURsi HeLa cells and incubated for 60 min, western blotting was performed to determine the phosphorylation of CHK1. (b) sicon and PLAURsi HeLa cells in the inserts, were treated with Dox (2 μM) for 1 h, and then placed in co-culture with untreated cells. Viability of the cells in the inserts was assessed after 6 days using CCK-8. Data shown as mean ±S.D. (c) HeLa cells pre-treated with TLR4 inhibitor (10 μg/ml) were stimulated with radiation (9 Gy) or Dox (2 μM), lysates were made after 2 and 4 h. Western blotting was performed for phosphorylation of CHK1. (d) HeLa cells irradiated at 9 Gy were fixed after 90 min and immunofluorescence was performed to detect colocalization of PLAUR and TLR4. Scale bar 40 μm