FIGURE 5.

Loss of Aerrie causes DNA damage and activation of signaling, while loss of YBX1/2/3 results in impaired DNA damage signaling. (A–C) HUVECs were treated with gapmeR (gap) targeting Aerrie, siRNA (si) targeting YBX1/2/3 or a respective control. (A) DNA damage was quantified by comet assay. HUVECs were stimulated with doxorubicin (Dox, 50 nM), embedded in low melting agarose, and stained with SYBR gold for imaging by fluorescence microscopy. Comets were quantified by CometScore (n > 3, at least 50 comets per experiment). (B) Phosphorylation levels of activated proteins in the double-strand break repair pathway were analyzed by western blotting. Phosphorylated ATM at serine 1981 and phosphorylated CHK-2 at tyrosine 68 were analyzed of Aerrie and YBX1 depleted HUVECs (n > 3). (C) Phosphorylation levels of activated proteins in the single-strand break repair pathway were analyzed by western blotting. Phosphorylated ATR at serine 1981 and CHK-1 at serine 317 were analyzed of Aerrie and YBX1 depleted HUVECs (n = 3). **p < 0.01; ****p < 0.0001; ns, not statistically significant.