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. 2014 Nov 6;42(21):13174–13185. doi: 10.1093/nar/gku1095

Figure 1.

Figure 1.

Hug1 level increases in response to DNA damage or replication blocks. (A) Exponentially growing cells were either treated with 0.1 M HU (black squares) or 0.05 μg/mL 4NQO (gray triangles) or mock treated (light gray squares). Relative mRNA levels were evaluated using the ACT1 gene as a standard. Analyses were performed at different time points as indicated. (B) Exponentially growing cells were either treated with 0.1 M HU or 0.05 μg/mL 4NQO or mock treated (no treatment). Total protein extracts were prepared at different time points as indicated during genotoxic treatment. Equal amounts of total protein extracts were loaded and analysed by SDS-PAGE followed by western blotting using anti-Hug1 serum. (C) Wild-type (WT) or RAD53-DL (DL) cells were grown in rich medium. Equal amounts of total protein extracts were loaded and analysed by SDS-PAGE followed by western blotting using polyclonal anti-Hug1 serum and anti-β-tubulin antibodies. (D) Exponentially growing wild-type cells were treated with 0.1 M HU for 2 h before release. Total protein extracts were prepared at different time points after release as indicated. Equal amounts of total protein extracts were loaded and analysed by SDS-PAGE followed by western blotting using polyclonal anti-Hug1 serum and monoclonal anti-GAPDH antibodies. Asyn: asynchronous cells.