Fig. 2: Loss of EWS-WT1 leads to morphologic changes and growth arrest in DSRCT cells.
From: Desmoplastic small round cell tumor is dependent on the EWS-WT1 transcription factor
a JN-DSCRT-1 cells undergo a morphologic change with EWS-WT1 silencing (siEWS-WT1) compared with untreated cells (Medium), a non-targeting siRNA (siNeg), or a positive control siRNA (siDeath). b, c Western blot demonstrates accumulation of the apoptotic marker cleaved PARP following EWS-WT1 silencing (b) that parallels cleaved caspase 3/7 (green cells) after EWS-WT1 silencing in live cells (c) occurring long after the morphologic change. Lysates were collected at 16, 24, 30, 48, 72, and 96 h of exposure. Western blot probed with EWSR1 (11910, Cell Signaling), H3 (2650, Cell Signaling), and cPARP (9546, Cell Signaling) and WT1 (sc-7385, Santa Cruz Biotechnology) antibodies. Apoptosis was quantitated by measuring activation of CellEvent caspase 3/7 green detection reagent (ThermoFisher Scientific) in 10,000 cells/well in a 96-well plate following siRNA silencing of EWS-WT1.
