Supplementary Figure 7: RT–PCR and immunoblotting for ERG isoforms.
From: Deregulation of DUX4 and ERG in acute lymphoblastic leukemia
(a) RT–PCR for ERG transcripts using primers specific for exons 1 and 10, showing amplification of internally deleted transcripts. (b) Immunoblotting using an antibody specific to the C terminus showing the aberrant ERG peptide fragment (approximately 28 kDa) in novel cases in the test cohort, but not in non-novel B- and T-lineage ALL cases. (c) Immunoblotting of lysates from HEK293T cells transfected with MSCV-IRES-GFP vector expressing ERG alt (lane 2), several DUX4/ERG samples with (lanes 3 and 4) or without (lanes 5–7) deletion, and non-DUX4/ERG ALL samples (lanes 8 and 9). The size of the protein expressed from the cloned ERGalt transcript is identical to that of the protein expressed in patients, confirming that this transcript encodes the aberrant C-terminal ERG protein fragment observed in leukemic cells.
