Fig. 2

NSC 19630 inhibits cellular proliferation in patient-derived cells. a C91PL cells were exposed to increasing amounts of the WRN helicase inhibitor NSC 19630 (0, 0.2, 2, and 20 μM). After 72 h, cells were stained with annexin V to determine the percentage of apoptosis. The figures include the percentage of cells in the four quarters: Q1, Q2, Q3, and Q4. Q3 included the live cells that are annexin V and PI negative. Q4 included early apoptotic cells, which are annexin V positive and PI negative. Q2 included cells in late apoptosis, which are both annexin V and PI positive. Finally, Q1 included necrotic cells, which are PI positive and annexin V negative. A dose-dependent effect was noted. Experiment was performed multiple times in duplicate. Representative results are shown in the final figures. b Normal resting PBMCs and C91PL were exposed to increasing amounts of the WRN helicase inhibitor NSC 19630 (0, 0.2, 2, and 20 μM). After 72 h, cells were stained with annexin V and survival cells were graphed. Experiment was performed in duplicate. P values were calculated comparing NSC-treated cells to DMSO control by using paired and two-tailed Student’s t test and indicated in the figure. c HTLV-1-transformed (MT-4, C8166, C91PL, 1186.94) and ATL-derived (ED, TL, ATL-25, ATL-43T, ATL-55T, LMY1, KK1, SO4, KOB) cell lines and normal resting PBMCs with increasing doses of NSC 19630 (0.2, 2, and 20 μM) show inhibition of cellular growth as measured by using cell count. Experiment was performed multiple times in duplicate. Representative results are shown in the final figures. d, e Patient-derived cell lines ATL-25 and LMY1 were treated with DMSO or 3 μM of NSC 19630 for 72 h and stained with crystal violet to test the anti-proliferative property of the WRN helicase inhibitor. The results were confirmed with XTT assay and cell counts. Experiment was performed multiple times in duplicate. Representative results were shown in the final figures. P values were calculated by using paired and two-tailed Student’s t test and indicated in the figure