HDM-2 inhibitors decrease RRM2 polypeptide expression. (A) REC-1, Granta-519, and JVM-2 wtp53 MCL cell lines were treated for 24 hours with either vehicle, 5μM MI-63, 5μM Nutlin, or 0.5μM doxorubicin (DOX) as a positive control for 24 hours. Qualitative PCR was performed to detect RRM2 and RRM2B mRNA levels, as well as β2M as a loading control, and transcripts were visualized by native gel electrophoresis. Lanes marked +RT received 1 μL of cDNA stock solution, whereas those labeled with 1:10 received 1 μL of 1:10 dilution of +RT. Representative images are shown in both panels of 1 of 3 independent experiments. (B) Protein levels of RRM2 and RRM2B, as well as β-actin as a loading control, were determined in lysates from MCL cells treated as described in panel A.
