Figure 4

MYC mRNA and protein levels are negatively affected by PRKDC gene suppression in cancer cell lines. A) Different lymphoma cell lines were treated with increasing concentrations of the PRKDC inhibitor, KU0060648, and a proteasome inhibitor, MG132. After a 4 h drug exposure time, c-MYC protein levels were analyzed via immunoblotting with an anti-MYC antibody. PRKDC and GADPH protein levels were also monitored with anti-PRKDC and anti-GADPH antibodies, respectively. B) Cells from A were analyzed by RT-PCR for relative c-MYC mRNA expression levels. C) The H82 cell line expressing inducible PRKDC shRNA was exposed to doxycycline for 3 days. c-MYC, PRKDC and GADPH protein levels were analyzed by immunoblotting with anti-MYC antibody, anti-PRKDC and anti-GADPH antibodies, respectively. Gene knockdown efficiency with independent shRNA clones against PRKDC was quantified by RT-PCR as was relative c-MYC mRNA expression levels in these cell lines.