Fig. 1

In vitro toxicity of epigenetic modulators is mediated through induction of apoptosis in epithelioid sarcoma and rhabdoid tumor cell lines. A Half-maximal inhibitory concentration (IC₅₀) of epigenetic agents after 72 h, based on confluence (IncuCyte®) or measured by MTS assay (*), determined at Day 10^a or at Day 6^c, in N ≥ 3 biological replicates (2 replicates^b); n.d. = not done. Western Blot analyses showing: B INI1 expression in A204, VAESBJ and GRU1, with maintained expression in GRU1 epithelioid sarcoma; C Biological activity of panobinostat (10xIC₅₀: 100 nM for A204, 250 nM for VAESBJ, GRU1) by induction of p21 and H4-acetylation; D Reduced ERK and AKT activity after 24, 48 and 72 h. E Induced apoptosis in a dose- and time-dependent manner measured by Annexin V- and PI-FACS analysis; graph presents means ± SEM of N = 3 replicates for 10/25 nM (IC₅₀), 50/125 nM (5xIC₅₀) or 100/250 nM (10xIC₅₀) for rhabdoid tumor/epithelioid sarcoma cell lines as compared to control (CO), and percentage of cells in early apoptosis (EA), late apoptosis (LA), or necrosis (N). F Confirmation of apoptosis by the presence of cleaved PARP and cleaved Caspase 3 in Western Blot