Truncated CCND1 leads to increased CCND1 mRNA, increased CCND1 protein expression, and cell-cycle progression. (A) Primer design for qRT-PCR to assess relative values of truncated versus full-length CCND1 mRNA. The cross exon primer set will amplify only mRNA in the coding region, thus avoiding genomic contamination. The proximal primer set will amplify both the full-length and truncated mRNA. The distal primer set will amplify only the full-length mRNA. (B) qRT-PCR showing the log ratio of truncated to full-length CCND1 mRNA in MCL cell lines. mRNA is expressed as relative quantitation equalized to beta-actin. Jeko-1 and Z138 had much higher quantitation values for truncated message and lower values for full-length message compared with Granta-519 and JVM2. Similarly, the ratio of truncated to full-length CCND1 mRNA is much higher in Jeko-1 and Z138. (C) Relative expression of total amount of CCND1 mRNA and corresponding CCND1 protein expression in 4 MCL cell lines. Cross exon primers were designed to amplify total CCND1 mRNA to avoid genomic contamination. Jeko-1 and Z138 also have increased total amount of CCND1 mRNA compared with Granta-519 and JVM-2. Similarly, Jeko-1 and Z-138 have increased CCND1 protein expression compared with Granta-519 and JVM-2. (D) Cell cycle in MCL cell lines. Jeko-1 and Z138 have increased percentage of cells in S-phase compared with Granta-519 and JVM-2.