(A) Western blot analysis over 3 hr of cleavage of CASPASE 3 (CASP3), total p53, p21, and c-MYC in a p53 wt wild-type Eμ-Myc lymphoma cell line treated with 50 nM CX-5461 in culture. See also Figure S4A.
(B) Relative p21, Puma, Odc1, and mTert mRNA expression determined by qRT-PCR (n = 3).
(C) Total RNA per cell (p > 0.05; n = 3) in cells from (a) at 1 hr of CX-5461 treatment.
(D) Nucleolar disruption shown in cells from (a) by fibrillarin (FBL) immunofluorescence with DAPI counterstain, nucleoli indicated by white arrows. See also Figure S4B.
(E) rDNA fluorescent in situ hybridization (FISH) with DAPI counterstain 2 hr post CX-5461 treatment. See also Figures S4C and S4D.
(F) Coimmunoprecipitation of ribosomal proteins L5 (RPL5) andL11(RPL11) with MDM2 (as comparedto GFP control)in vehicle-treated Eμ-Myc lymphoma cells overexpressing BCL2 cells versus cells treated with 500 nM CX-5461, as shown by western immunoblot (IB) analysis. Images taken at 60× magnification. Scale bar represents 10 μm. Student's two-tailed t test for all comparisons. Error bars represent SEM. See also Figure S4D.