Figure 3 from Histone Deacetylase Inhibitors Target DNA Replication Regulators and Replication Stress in Ewing Sarcoma Cells
Year: 2025
Type: other
Abstract: HDAC inhibitors impair DNA replication in Ewing sarcoma cells. A, Representative cell-cycle analysis (EdU and propidium iodide) plot for EW8 cells treated with romidepsin (5 nmol/L) for 24 hours. B, Summary of EdU incorporation efficiency for additional Ewing sarcoma cell lines treated with romidepsin (5 nmol/L) for 24 hours. C, Ewing sarcoma cell lines were treated with romidepsin (5 nmol/L) for 24 hours, and then chromatin-bound RPA2 was assessed using immunoblotting. D, EW8 cells expressing a doxycycline-inducible shRNA targeting RPA2 were treated with doxycycline for 24 hours, and then cellular lysates were collected for immunoblotting. E and F, EW8 cells expressing a doxycycline-inducible shRNA targeting RPA2 were treated with doxycycline (1,000 ng/mL) for 24 hours, and then dose-response experiments were performed with romidepsin (E) and panobinostat (F). Cell viability was measured 72 hours after drug addition using the alamarBlue assay. Error bars represent the mean ± SD of three technical replicates. The results are representative of two independent experiments. G, CRISPR dependency score for CDT1 and MCM2–7 (depmap.org). H, EW8 and TC71 cells were treated with two different siRNAs targeting MCM2, and then cellular lysates were collected for immunoblotting. I, EW8 and TC71 cells were treated with siRNAs targeting MCM2, and then EdU incorporation into DNA was measured using flow cytometry. J, Kaplan–Meier analysis shows overall survival of patients with Ewing sarcoma tumors according to the expression level of CDT1 and MCM2 mRNA. P values were calculated using a two-tailed Student t test or a one-way ANOVA followed by the Dunnett multiple comparisons test. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4411705114