FIGURE 2 from Activity of the Ubiquitin-activating Enzyme Inhibitor TAK-243 in Adrenocortical Carcinoma Cell Lines, Patient-derived Organoids, and Murine Xenografts

Cytotoxicity of TAK-243 on ACC cell lines. A, Concentration–response curve and IC₅₀ values of TAK-243 in ACC cell lines and SW-13. Cell viability after 72 hours under the indicated drug concentrations was measured by the CellTiter-Glo viability assay. B, Expression of ABC transporters (MDR-1, BCRP) and SLFN11 in ACC cell lines and SW-13. Proteins were extracted from each cell line and expression of SLFN11, MDR-1, and BCRP were evaluated by Western blotting. C, Effects of TAK-243 on DNA synthesis and cell cycle. CU-ACC1, CU-ACC2, and NCI-H295R cells were treated with 100 nmol/L TAK-243 for the indicated times and pulse-labeled with EdU (10 µmol/L) for 30 minutes before harvest. Cellular EdU uptake and cell cycle were analyzed by flow cytometry. D, Change in percentage of S-phase after TAK-243 treatment. E, ACC cell lines and SW-13 were treated with TAK-243 at the indicated concentrations for 24 hours, and PARP cleavage was detected by Western blotting.