Research Paper Volume 12, Issue 5 pp 4547—4557

Figure 2. ARV-825 induces apoptosis activation in human thyroid carcinoma cells. TPC-1 cells (A–D), the primary human thyroid carcinoma cell (“C1”/“C2”, G and H) or the primary human thyroid epithelial cells (“E1”/“E2”, G and H) were treated with ARV-825 (5-250 nM) and cultured in for indicated time periods, the caspase activity (A), expression of apoptosis-associated proteins (B) as well as nuclear TUNEL staining (C and G) and mitochondrial depolarization (JC-1 green fluorescence intensity, D and H) were tested to examine cell apoptosis. TPC-1 cells were pre-treated for 30 min with 50 μM of z-DEVD-fmk, z-LEHD-fmk or z-VAD-fmk, following by ARV-825 (100 nM) treatment for 48-72h, cell apoptosis and cell viability were tested by TUNEL staining (E) and MTT assay (F), respectively. Expression of the listed proteins was quantified and normalized to loading control (B). “DMSO” stands for vehicle control (0.1% DMSO, E and F). **p < 0.01 vs. “Ctrl” group. ***p < 0.001 vs. “Ctrl” group. ^# p < 0.001 vs. “DMSO” group (E and F). The experiments were repeated three times, with similar results obtained. Bar=100 μm (D).